Structural analogues of vitamin D

ABSTRACT

A compound of the formula I:                    
     in which 
     P is hydrogen or alkyl; 
     Y and Y′ are each hydrogen or, when taken together, represent a group ═CH 2 ; 
     W and W′ are each hydrogen; 
     X is selected from the group consisting of a hydroxyalkyl, a hydroxyalkoxy, an alkoxy optionally comprising an epoxide function, a hydroxyalkene, a hydroxyalkadiene, a hydroxyalkyne and isomeric forms thereof; and 
     a) either R 1  and R 3  or R′ 3  form a saturated 5- or 6-membered carbocyclic ring and R 2 , R′ 2 , R 3  or R′ 3 , R 4 , R′ 4 , R 5  and R′ 5  are each independently hydrogen or alkyl; or 
     b) R 2  or R′ 2  and R 4  or R′ 4  form a saturated 5- or 6-membered carbocyclic ring, and R 1 , R 2  or R′ 2 , R 3 , R′ 3 , R 4  or R′ 4 , R 5  and R′ 5  are each independently hydrogen or alkyl; or 
     c) R 3  or R′ 3  and R 5  or R′ 5  form a saturated or unsaturated 5- or 6-membered carbocyclic ring, and R 1 , R 2 , R′ 2 , R 3  or R′ 3 , R 4 , R′ 4 , and R 5  or R′ 5  are each independently hydrogen or alkyl; or 
     d) R 3  or R′ 3  taken at the same time with R 1  and R 5  or R′ 5  form a saturated 8- or 12-membered carbobicyclic ring, and R 2 , R′ 2 , R 3  or R′ 3 , R 4 , R′ 4 , and R 5  or R′ 5  are each independently hydrogen or alkyl.

The present application is a continuation of U.S. application Ser. No.08/571,887, filed Jun. 28, 1996, now U.S. Pat. No. 6,017,907 which is a371 of PCT/EP94/02294 filed Jul. 7, 1994.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention describes a hitherto unknown and therefore new class ofcompounds which are analogues of 1α,25-(OH)₂D₃ and show selectiveactivity on cell functions.

2. Description of Related Art

General Introduction

Vitamin D of either nutritional (vitamin D₂ or D₃) origin or produced inthe skin under the influence of ultraviolet light is metabolized inseveral tissues to produce firstly 25-hydroxyvitamin D₃ [25-OHD₃] andlater 1α,25-dihydroxyvitamin D₃ [1α,25-(OH)₂D₃] and numerous othervitamin D metabolites (1-6). Several hydroxylases present in differenttissues (e.g. liver, kidney, placenta, keratinocytes, fibroblasts,monocytes, lymphocytes, bone cells . . . ) are responsible for both theactivating and inactivating pathways of the parent vitamin D molecules.1α,25-(OH)₂D₃ behaves as a classical steroid hormone as its synthesis isfeedback controlled by several hormones, ions and humoral factors tomaintain a normal body homeostasis of plasma and bone minerals. Moreoverthe vitamin D hormone(s) act via binding and activation of specificvitamin D receptors, present in most tissues and cells. Thesteroid-receptor complex then functions as a transactivating factor bybinding to specific DNA sequences known as vitamin D responsive elementsso that transcription of numerous genes is either activated orinactivated (7,8). This gene (in)activation occurs in collaboration withother nuclear accessory factor(s) of which the vitamin A receptor (RXR)is part of (9,10). Moreover there is some evidence for the activity ofvitamin D, its metabolites and analogues to act via nongenomicmechanisms, either by activating calcium channels or other membrane orsecond messenger signals (11-13). Vitamin D, its metabolites andanalogues have potent effects on calcium and phosphate metabolism, andtherefore they can be used for prevention and therapy of vitamin Ddeficiency and other disorders of plasma and bone mineral homeostasis(e.g. osteomalacia, osteoporosis, renal osteodystrophy, disorders of theparathyroid function). Moreover vitamin D receptors are found innumerous tissues and cells that do not belong to the target tissuesresponsible for the just mentioned calcium homeostasis. Such cellsinclude most cells belonging to the endocrine system and vitamin D, itsmetabolites and analogues are capable of influencing the hormonalsecretion of these glands or tissues (e.g. insulin, parathyroid,calcitonin, pituitary hormones). Vitamin D receptors and vitamin Dactivity have also been documented in calcium transporting tissues otherthan the intestine and bone (e.g. placenta and mammary glands). Inaddition vitamin D receptors and vitamin D action have been observed inmost other cells (e.g. cells belonging to the immune system, skincells). These cells or tissues can be of a benign, adenomatous or of amalignant type. These so-called noncalcemic effects of vitamin D, itsmetabolites and analogues create the possibility to use such compoundsfor various therapeutic applications such as modification of the immunesystem, modification of hormone secretion, altering calcium transport inseveral tissues, modification of intracellular calcium concentration,induction of cell differentiation or inhibition of cell proliferation(14,15). In particular such compounds may be useful in the therapy ofdisorders characterized by increased cell proliferation (e.g. psoriasis,cancer) (16-18).

To increase the therapeutic potential of the natural vitamin Dhormone(s), analogues can be synthesized with increased potency for aspecific action and reduction of another type of action. For example toobtain an anti-psoriasis drug analogues can be synthesized with anincreased activity on keratinocytes and lymphocytes present in theaffected skin areas but with decreased effects on serum, urinary or bonecalcium (19-23). Similarly analogues can have an increased potency toinhibit proliferation of cancer cells (e.g. leukemia or breast cancercells) and/or increase the differentiation of such cells, either aloneby their intrinsic potency or enhance such effects in combination withother drugs (e.g. growth factors or cytokines, other steroid orantisteroid hormones or retinoic acids or related compounds) and at thesame time have a reduced potency to influence serum, urinary or bonecalcium or phosphate homeostasis. Another such example would beanalogues with increased activity on specific hormone secretion (e.g.parathyroid hormone, insulin) without the same relative potency for theother activities of the natural vitamin D hormone(s). Analogues withincreased activity on non-malignant cells belonging to the immune systemcould be used for the treatment of immune disorders (e.g. autoimmnunedisorders, AIDS, prevention of graft rejection or graft versus hostreaction) especially if their effect on other systems (e.g. calcium andphosphate metabolism) would be relatively weakened. Moreover analoguescan be developed with increased activity on bone forming cells without asimultaneous potency on bone resorbing cells or vice versa and suchanalogues could be useful in the treatment of bone disorders.

A number of vitamin D analogues with modifications in the specificaction in different tissues (especially the potency ratio on celldifferentiation and calcemic effects) have been described previouslywith variable success in such differentiation. Especially oxa analoguesin the side chain (patent WO 90/09992; EP 0385 446A 2), modifications orhomologation of the side chain (WO 87/00834, international patentclassification CO7C 172/00), changes in the stereochemistry at carbon 20(WO 90/09991, international patent classification CO7C 401/00, A61K31/59), modifications on C11 of the C ring (EP 89/401,262-4) and epoxyanalogues (PCT/EP 92/0126) of the side chain displayed interestingcharacteristics.

SUMMARY OF THE INVENTION

Present Invention

The present invention relates to the synthesis and biological evaluationof original compounds which still maintain some of the essentialcharacteristics of vitamin D action but with a more selective pattern,(i.e. not all the actions of the physiological vitamin D hormone aremaintained with the same relative potency) and with a structure that canbe thoroughly modified in the central part. Indeed within the structureof vitamin D one may distinguish three different parts : (i) a centralpart consisting of the bicyclic CD-ring system; (ii) an upper part,consisting of the side-chain which is connected to position 17 of theD-ring; (iii) a lower part, consisting of the A-ring and theΔ(5,7)-diene (the so-called seco B-ring), which is connected to position8 of the C-ring. One aim of the present invention is to bring aboutsubstantial structural modifications in the central part of vitamin D.

In particular the present invention relates to analogues of vitamin D,which lack the combined presence of the trans-fused six-membered C-ringand of five-membered D-ring, but still possess a central part consistingof a substituted chain of five atoms, atoms which correspond topositions 8, 14, 13, 17 and 20 of vitamin D, and at the ends of whichare connected, at position 20 a structural moiety representing part ofthe side-chain of vitamin D or of an analogue of vitamin D, and atposition 8 the Δ(5,7)-diene moiety connected to the A-ring of the active1-alpha-hydroxy metabolite or of an established vitamin D analogue.

BRIEF DESCRIPTION OF THE DRAWINGS

In the accompanying drawings, the legends correspond to those which areshown in FIGS. 1A and 1B.

FIGS. 1A and 1B are graphs of affinity for the vitamin D receptor forpig intestinal mucosa for selected vitamin D analogues;

FIGS. 1C and 1D are graphs of affinity for the vitamin D receptor forhuman vitamin D binding protein for selected vitamin D analogues;

FIGS. 2A and 2B are graphs of induction of cell differentiation in humanpromyeloid leukemia cells (HL-60) by selected vitamin D analogues asevaluated by potency to induce superoxide production measured by NBTreduction;

FIGS. 3A and 3B are graphs of induction of cell differentiation in humanosteosarcoma cells (MG-63) by selected vitamin D analogues as evaluatedby potency to induce ostecalcin secretion;

FIG 3C is a graph of induction of cell differentiation in humanosteosarcoma cells (MG-63) by selected vitamin D analogues as evaluatedby potency to inhibit cell proliferation in such cells as measured by[³H]thymidine incorporation;

FIGS. 4A-4C are graphs of inhibition of cell proliferation bynonsteroidal vitamin D analogues in human breast cancer cells (MFM-223and MCF-7) by selected vitamin D analogues as evaluated by [³H]thymidineincorporation;

FIGS. 5A and 5B are graphs of inhibition of human keratinocyte cellproliferation by selected vitamin D analogues as evaluated by potency toinhibit [³H]thymidine incorporation in vitro; and

FIGS. 6A-6D are graphs of calcemic effects of selected vitamin Danalogues as evaluated in vitamin D deficient chicks after i.m.treatment for ten consecutive days, with serum and bone calcium serumosteocalcin and duodenal calbindin concentration measured to evaluatethe calcemic potency.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of the invention are represented by the general formula I,in which formula:

P stands for hydrogen, alkyl or acyl;

X represents part of the side-chain of vitamin D or of one of itsestablished analogues;

Y and Y′, which may be the same or different, stand for hydrogen oralkyl or, when taken together, represent an alkylidene group, or form acarbocyclic ring;

W and W′, which may be the same or different, stand for hydrogen oralkyl or, when taken together, represent an alkylidene group, or form acarbocyclic ring;

one of the carbon atoms of the central part corresponding to positions14, 13, 17 or 20, together with the R and R′ substituents connected toit, may be replaced by an oxygen (O), a sulfur (S) or a nitrogen bearingan R substituent (NR).

R and R′ (i.e., R, R₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅):

when located in a relative 1,3-position along the central chain, such asR₁ and R₃ or R′₃, R₂ or R′₂ and R₄ or R′₄, R₃ or R′₃ and R₅ or R′₅,taken together with three adjacent atoms of the central chain, whichcorrespond to positions 8, 14, 13 or 14, 13, 17 or 13, 17, 20,respectively, can form a saturated or unsaturated carbocyclic orheterocyclic 3-, 4-, 5-, 6- or 7-membered ring also including caseswhereby geminal substituted R and R′ taken together form a cyclicunsaturated bond, under the proviso that when R₁ and R′₃ form a6-membered carbocyclic ring of the following nature (1) unsubstitutedand saturated (2) monosubstituted at C-11 or (3) having a double bondbetween C-9 and C-11, R₂ and R₄ do not form a five-membered carbocyclicring when R₃ is methyl, etnyl or ethenyl.

when located in a relative 1,2-position (i.e., vicinal) along thecentral chain, such as R₁ and R₂ or R′₂, R₂ or R′₂ and R₃ or R′₃, R₃ orR′₃ and and R₄ or R′₄, R₄ or R′₄ and R₅ or R′₅, and when not being partof a ring as described above, taken together with two adjacent atoms ofthe central chain, which correspond to positions 8,14 or 14,13 or 13,17or 17,20, respectively, can form a saturated or unsaturated carbocyclicor heterocyclic 3-, 4-, 5-, 6- or 7-membered ring, also including caseswhereby geminal substituted R and R′ taken together form a cyclicunsaturated bond.

when located in a relative 1,1-position (i.e., geminal) along thecentral chain, such as R₂ and R′₂, or R₃ and R′₃, or R₄ and R′₄ or R₅and R′₅, and when not being part of a ring as described above, takentogether with the carbon bearing the R and R′ substituents can formeither a saturated or unsaturated carbocyclic or heterocyclic 3-, 4-,5-, or 6-membered ring.

which may be the same or different, and when they are not forming a ringor a bond as described above, stand for hydrogen or a lower alkyl group,or when taken together in the case of geminal substitution represent alower alkylidene group.

In the context of the invention the expression “lower alkyl group”indicates a straight or branched saturated or unsaturated carbon chaincontaining from 1 to 7 carbon atoms, and “lower alkylidene group”indicates a straight or branched saturated or unsaturated carbon chaincontaining from 1 to 7 carbon atoms, which is connected to one of themain chain atoms 14, 13, 17 and/or 20 through a double bond.

In the context of the invention part of the side-chain of vitamin D orof one of its established analogues stands for a 2 to 15 carbon atomsubstituted alkyl chain especially as present in vitamin D₂ (C-22 toC-28) or D3 (C-22 to C-27) or partially modified as shown below with thevitamin D numbering, especially:

hydroxyl substituent at one or more positions, for instance 24, 25and/or 26 and/or

methyl or ethyl substituent in one or more positions, for instance 24,26 and/or 27 and/or

halogen substituent(s) at one or more positions for instanceperfluorated at positions 26 and/or 27 or difluorated at position 24and/or

additional carbon atom(s) especially C₂₄ between the positions 24 and25, with the same substitution pattern mentioned above and/or

esters derivatives of one or more hydroxyl substituents mentioned aboveand/or

changing one or more carbon atoms for an oxygen, nitrogen or sulfur atomfor instance at the positions 22, 23 or 24 and/or

cyclized between the carbon atoms 26 and 27 by one bond (cyclopropane)or by the intermediacy of 1 to 4 carbon atoms, the ring can besaturated, unsaturated or aromatic and may optionally be substituted atany possible position(s) with the substituent mentioned above and/or

cyclized between the carbon atoms 26 and 27 by 1 to 4 atoms to form aheterocyclic ring, including aromatic, which may optionally besubstituted at any possible position with the substituent mentionedabove and/or

unsaturated with one or more double or triple C—C bond(s), theseunsaturated chains may be substituted at any possible position by thesubstituents mentioned above and/or

epoxide function can be present between carbon atoms 22,23 or 23,24 or24,25 or 25,26; these epoxidized chains can be saturated or unsaturatedand may be substituted at any possible positions with the substituentsmentioned above and/or

two or more of the carbon atoms of the side chain can be linked by asingle bond or by the intermediacy of a one to five carbon or oxygen,nitrogen or sulfur atoms to form a 3-7 membered saturated or unsaturatedcarbocyclic or heterocyclic including aromatic ring which may optimallybe substiuted at any possible position by substituents mentioned aboveand/or

substituted at one or more positions by saturated, unsaturatedcarbocyclic, heterocyclic or aromatic ring which can be substituted atany possible position(s) with the substituents mentioned above

isomeric forms of the substituted chain.

Hence the invention relates to a series of analogues with widely varyingstructures as exemplified in Table 1 where some specific examples ofcompounds with formula I are shown and which are referred to by numberin the preparations and examples.

Most often the compounds of the invention are represented by one of theformulas IIa (type C), IIb (type D), IIc (type E), IId (type CD), IIe(type CE), IIf (type DE), and IIg (acyclic type):

where:

X, Y, Y′, W and W′ have the same meaning as above;

Z represents a saturated or unsaturated hydrocarbon chain consisting ofzero (hence Z represents a bond between two 1,3-related carbon atoms ofthe central chain), one, two, three or four atoms, which may all besubstituted and/or replaced by a heteroatom such as oxygen, sulfur andnitrogen.

R₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅

which may be the same or different, stand for hydrogen or lower alkyl,such as methyl, ethyl or n-propyl.

Among those are preferred the cyclic derivatives of type C, D, E, CD, CEand DE that correspond to structures IIIa, IIIb, IIIc, IIId, IIIe, andIIIf, respectively,

wherein

n is an integer equal to 2 or 3

X represents one of the following vitamin D side-chain parts(4-hydroxy-4-methyl)pentyl, (R)- or (S)-(3-hydroxy-4-methyl)pentyl,(3′-hydroxy-3′-methyl)butyloxy, (4-hydroxy-4-ethyl)hexyl,(4-hydroxy-4-methyl)-2-pentynyl, (4′-hydroxy-4′-ethyl)hexyloxy;4,5-epoxy, 4-methyl-2-pentynyl; 4-hydroxy-4-ethyl-2-hexynyl;(3-methyl-2,3-epoxy)-butyloxy; (3-hydroxy-3-ethyl)-pentyloxy;(4-hydroxy-4-ethyl)-hexyloxy

Y, Y′, W and W′ are the same and represent hydrogen, or taken togetherrepresent a methylene group ═CH₂;

R₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅ and R′₅, which may be the same ordifferent, stand for hydrogen or methyl.

All compounds of the invention can be prepared using reactions that arewell-known in the art of synthetic organic chemistry. In particular, inall cases, the lower part of the structure can be introduced followingthe method of Lythgoe (24) whereby the anion of a protected phosphineoxide IV is reacted with the appropriate carbonyl derivative VII, inwhich the various reactive functional groups are preferentiallyprotected and in which the groups X, Y, Y′, W, W′, Z, P, R₁, R₂, . . .R′₅ have the same meaning as previously, whereafter the reactivefunctional groups are deprotected. Also, the synthesis of derivatives asIV has been reported in the literature (25).

Alternative constructions involve (a) coupling of an appropriate vinyliccarbanion (from VIII) with V followed by acid catalyzed solvolysis and(b) reaction of the alkynyl anion of VI with an appropriate carbonylderivative VII followed by partial triple bond reduction and acidcatalyzed solvolysis (26). It should also be possible to adapt the routeso that alternative coupling methods can be used such as the sulfone way(27a) or Okamura's coupling (27b).

The compounds with structure VII can be obtained following variousroutes as will be shown with several examples. It is important to notethat those derivatives will generally be obtained following syntheticroutes that are shorter and more efficient than those that are usuallyused in the preparation of analogues of vitamin D.

(a) AlCl₃, isoprene, toluene, 6 h, −78° C.—r.t. (72%); (b) MeONa, MeOH,1 h, r.t. (94%); (c) NaBH₄, MeOH, 12 h, 0° C.—r.t. (86%); (d) MEMCl,DIPEA, THF, 3 h, r.t. (98%); (e) (i) OsO₄, NMMO, Me₂CO:H₂O (3:1), 12 h,r.t. (86%); (ii) NalO₄, Me₂CO:H₂O (3:1), 12 h, r.t. (98%); (f) KOH, 12h, 60° C. (53%); (g) (i) 10% Pd/C, 1 atm H₂, hexane, 1.5 h, 0° C. (93%);(ii) MeONa, MeOH, 3 h, 0° C.—r.t. (97%); (h) Ph₃P═CH₂, HMPA:THF (1:1), 2h, −20° C. (100%); (i) (i) 9-BBN, THF, 4 h, r.t.; (ii) EtOH, NaOH 6N,H₂O₂ 30%, 1 h, 60° C. (74%-91%); (j) TsCl, DMAP, Et₃N, CH₂Cl₂, 12 h,r.t. (91%-97%); (k) NaH, DMSO, 2-(1-ethoxy)-ethyloxy-2-methyl-3-butyne,1.5 h. 60° C.—r.t. (70%); (l) 10% Pd/C, 4 bar H₂, EtOAc, 1 h, r.t.(34%); (m) Me₂BBr, CH₂Cl₂, 1 h, −78° C. (73%); (n) PDC, CH₂Cl₂, 4 h,r.t. (86%-99%); (o) TSIM, THF, 1 h, r.t. (94%-98%); (p) TBAF, THF, 5days, 30° C. (99%); (q) (i) NaH, CS₂, 24 h, r.t.; (ii) Mel, THF, 2 h,r.t. (98%); (r) Bu₃SnH, AIBN, toluene, 9 h, 110° C. (92%); (s) KI, DMSO,4 h, 60° C. (95%); (t) methylvinylketone, Cul, Zn, EtOH:H₂O (7:3), 3.15h, 15° C. (83%); (u) MeMgCl, THF, 1 h, r.t. (98%); (v) Amberlyst 15,MeOH, 1 week, 30° C. (96%).

Scheme 1

The 18-nor-vitamin D skeleton is a representative of analogues of typeIIId. The synthesis centers around the following steps: (a) synthesis ofa transfused decalone, (b) one-ring contraction to a trans-fusedhydrindane, (c) side chain construction.

The, in the literature described dienophile 1.1, is known to giveDiels-Alder addition syn to the silyloxy group (28). Thus,regioselective reaction with isoprene gives 1.2a; base inducedepimerization leads to 1.2b. Selective reduction of the carbonylfunction and subsequent alcohol protection leads to intermediate 1.3b.Double bond cleavage and aldol reaction of the resulting dialdehydegives trans-hydrindane 1.4. Hydrogenation of 1.4 leads to a mixture ofC-17 epimers which is, upon base induced epimerization, transformed intothe thermodynamically more stable 1.5a. Wittig reaction andhydroboration leads to 1.6a next to circa 20% of the C-20 epimer. Afterseparation, the side chain is introduced via tosylate 1.6b. Finallly,catalytic hydrogenation, reintroduction of the C-8 carbonyl function and25-hydroxyl protection afford the desired precursor 1.8d. Intermediate1.5b also allows the removal of the C-12 oxy-function via a wellestablished procedure, involving a radical reaction (29).

Hydroboration of 1.9c and subsequent transformation of the hydroxylgroup into iodo-compound 1.10c (4:1, 20S:20R). The side chain isintroduced under sonication conditions yielding 1.10d (30). This ketonegives, upon reaction with methyl magnesiumchloride, the tertiary alcohol1.11a. Oxidation to the C-8 ketone 1.11.c and tertiary alcoholprotection affords the desired precursor 1.11d.

Analogues with the six-membered structure IIIa can be synthesizedaccording to a strategy which involves as a key-step the Ireland-Claisenrearrangement of a substrate obtained from an ester of which the alcoholpart consists of (R)-3-methyl-2-cyclohexenol (31). Two examples of thisstrategy are shown in scheme 2.

Reaction of (R)-3-methyl-2-cyclohexenol with the homochiral acid 2.1obtainable from (−)-menthone (32), gives the ester 2.2. Afterdeprotonation of the ester, the enolate anion is reacted in situ withtert-butyldimethylsilyl chloride; subsequent thermolysis leads tocyclohexene 2.3 (67% after recovery of starting material) (33). Thecarboxygroup in 2.3 is subsequently transformed into a methylgroup,following standard conditions, yielding eventually derivative 2.4.Hydroboration of 2.4 gave a secondary alcohol which is oxidized tocyclohexanone 2.5. The latter is the required carbonyl substrate for thesynthesis of analogues 4 possessing the (24S)-configuration.

(a) DCC, DMAP, CH₂Cl₂ (91%); (b) LiCA, THF, HMPA; tBuMe₂SiCl; (c) Δ(67%); (d) CH₂N₂, ether (86%); (e) LAH, THF (89%); (f) TsCl, Pyridine(96%); (g) LAH, THF (91%); (h) 9-BBN, THF; NaOH, H₂O₂ (80%); (i) PDC,CH₂Cl₂ (90%); (j) TBAF, THF, 30° C. (88%); (k) PPh₃, DEAD, pNO₂PhCOOH(68%); (l) K₂CO₃, KOH; (m) TBSCl, imidazole, DMF, DMAP (97%); (n) 9-BBN,THF (92%); (o) PDC, CH₂Cl₂ (92%); (p) DCC (96%); (q) LDA, TBSCl; (r)LAH, THF, Δ (86%); (s) TsCl, py (100%); (t) LAH, THF (100%); (u)Hg(OAc)₂, NaOH, NaBH₄; (v) TESCl, DMAP, DMF, imidazole; (w) 9-BBN, H₂O₂(95%); (x) PDC (80%).

Scheme 2

The synthesis of its (24R)-epimer is performed in a similar way afterinversion at C-24. Therefore starting from intermediate 2.4, theprotective group is removed and the resulting alcohol inverted via theMitsunobu procedure (34). Repetition of the same sequence as above givescyclohexanone 2.7. The usual coupling procedure then leads eventually toanalogues 5 and 6 which possess the (24R)-hydroxy group.

The synthesis of the 25-hydroxy analogue can be performed along the samestrategy. Therefore (R)-3-methyl-2-cyclohexenol is esterified with(R)-(+)-citronellic acid (2.8) to yield ester 2.9. The Ireland-Claisenrearrangement sequence then gives the acid 2.10. After transformation ofthe carboxygroup into a methyl group (2.11), the trisubstituted doublebond is preferentially oxidized to the tertiary alcohol using mercuricacetate, NaOH and sodium borohydride. Subsequent alcohol protection andregioselective oxidation of the cyclic double bond leads tocyclohexanone 2.12, from which are obtained, using the usual couplingprocedure, analogues 7 and 8.

Analogues of type IIIa with inverted configuration at C-13 can also beobtained via Ireland-Claisen strategy. This is illustrated in scheme 3.For that purpose the acetate of (S)-3-methyl-2-cyclohexenol (3.1; 86%ee) can be directly deprotonated, and the corresponding enol silyletherrearranged to the acid 3.2. A further enrichment of the desiredenantiomer can be realized via resolution with R-(+)-α-methylbenzylamine. The subsequent sequence involves reduction of acid 3.2, andprotection of the resulting primary alcohol to 3.3. The latter can beoxidized using 9-BBN and hydrogen peroxide to alcohol 3.4. Afterprotection-deprotection, the primary alcohol is used to build up an oxaside-chain. This is performed by reaction of the anion with1-chloro-3-methyl-2-butene. After hydrolysis and oxidation cyclohexanone3.6 is obtained. The final introduction of the 25-hydroxy group involvesthe mercuric acetate-hydride reduction method. The obtainedcarbonylderivative 3.7 serves as a precursor for analogue 9characterized by a 22-oxa sidechain and an epimeric configuration atC-13. One can further note that the usual Horner-Wittig coupling alsoleads in this case to the formation of the isomer with a (Z)-7,8-doublebond (ratio 4:1).

P=SiPh₂tBu

(a) PGL, phosphate buffer (86% ee); (b) LDA, tBuMe₂SiCl, THF; HCl;resolution with R-(+)-α-methyl benzylamine (48%); (c) LAH, ether (95%);(d) tBuPh₂SiCl, DMF, imidazole (98%); (e) 9-BBN, H₂O₂ (96%); (f) DHP,CH₂Cl₂ (93%); (g) (nBu)₄NF, THF (91%); (h) ClCH₂CH═C(CH₃)₂, NaH, DMF(81%); (i) TsOH, MeOH, r.t. (98%); (j) PDC, CH₂Cl₂, r.t. (84%); (k)Hg(OAc)₂, NaBH₄ (68%).

Scheme 3

Another strategy towards the synthesis of analogues of type IIIaconsists in the conjugate addition of part of the side chain involving3-methyl-2-cyclohexenone as the substrate. An example is given in scheme4.

(a) tBuPh₂SiCl, imidazole, DMF, 36 h, r.t. (100%; (b) DIBALH, hexane,0.5 h, −78° C.; (c) tBuOK, (MeO)₂P(O)CHN₂, THF, 20 h, −78° C., r.t. (90%overall from 4.2); (d) B—Br-9-BBN, CH₂Cl₂, 4 h, 0° C., then CH₃COOH, 0.5h, 0° C., NaOH/H₂O₂, 0.5 h, r.t. (90%); (e) tBuLi, Cul/HMPT, BF₃—OEt₂,3-methyl cyclohexenone, ether, 16 h, −120°-20° C. (40%); (f) TBAF, THF,3 h, r.t. (90%); (g) HPLC, eluent:hexane:ethylacetate 6:4; (h) Ph₃P,imidazole, I₂, THF, 6 h, −20° C.—r.t. (88%);

Scheme 4

The necessary homochiral cuprate reagent is obtained following asequence starting from methyl (S)-3-hydroxy-2-methylpropanoate (4.1).After protection of the alcohol, the ester is reduced and the resultingaldehyde 4.3 treated with the anion derived from methyl diazomethylphosphonate (35). The resulting alkyne 4.4, obtained in 90% yield from4.2, is subsequently transformed into the vinyl bromide derivative 4.5.From the latter an appropriate cuprate reagent is obtained via treatmentwith tert-butyllithium and Cul at −120° C. The 1,4-addition to3-methyl-2-cyclohexenone is performed in ether in the presence ofborontrifluoride (36). After usual work-up and purificationcyclohexanone 4.6 is obtained together with its C13-epimer.

After hydrolysis the desired alcohol 4.7 can be separated from itsC13-epimer (configurational assignment according to CD), and is furthertransformed into iodide 4.8. This carbonyl derivative serves as thesubstrate for appending the A-ring.

For the synthesis of compounds of type IIc, an example is given in.scheme 5. The starting material 5.1 is available from R-citramalic acid(37).

(a) TsOH, THF, 20 h, r.t. (90%); (b) DDQ, 3 h, r.t.; (c) PDC, DMF, 20 h,r.t.; (d) CH₂N₂, Et₂O (94%); (e) EtMgBr, 2 h, r.t.; (f) Pd/C, H₂ (50%);(g) TPAP, NMMO, 2 h, r.t. (70%);

Scheme 5

Construction of the heterocyclic nucleus from 5.1 and 5.2 allowsassembling of the precursor skeleton in a convergent way. Both epimersof 5.3 with respectively α and β oriented side chain are obtained in a1:1 ratio.

Further transformations are carried out on this epimeric mixture.Separation was possible on the stage of the final analogues.Transformation of the p-methoxybenzylether in 5.3 (α+β) into the ester5.4 (α+β) and subsequent Grignard reaction leads to the side chain.Finally the aldehyde function was introduced and affords the precursor5.6(α+β).

A group of analogues with a five-membered ring, as examples of thegeneral formula IIIc, can readily be obtained starting from the known6.1 (38). Cleavage of the ether bond in 6.1 with sodium iodide leads tothe key-intermediate, the iodide 6.2. The synthesis centers aroundintroduction of (a) the side chain using the iodo-function via (1)direct coupling or (2) after transforming the iodomethyl substituent toa hydroxyl substituent or (3) after inverting the orientation of theiodomethyl substituent or (4) after transformation of the iodide into aformyl group and of (b) the A-ring part after homologation at thehydroxymethyl substituent. Examples of this strategy are given below andare illustrated in scheme 6.

The iodo-compound 6.2 can be coupled under sonication conditions withmethyl-vinyl ketone and ethyl-vinyl ketone to yield respectively 6.8 and6.9. Ketone 6.8 upon reaction with methyl magnesiumbromide gives thecorresponding tertiary alcohol. Oxidation of the primary alcohol and 1-Chomologation of the resulting aldehyde 6.10 withmethoxy-triphenylphosphonium-methylide and subsequent hydrolysis leadsto the aldehyde 6.12 required for coupling with the A ring. Similarly,reaction of 6.9 with ethyl magnesium bromide and subsequenttransformation gave 6.13.

(a) Cl₃SiCH₃, NaI, CH₃CN (90%); (b) DIPEA, CH₃OCH₂Cl, CH₂Cl₂ (86%); (c)TBAF, THF (88%); (d) OsO₄, NaIO₄, THF:H₂O (65%); (e) LiAlH₄, THF, rt(95%); (f) (1) 9-BBN, THF, 60° C.; (2) H₂O₂, NaOH (87%); (g) Ph₃P,imidazol, I₂, ether:CH3CN 3:1 (93%); (h) Amberlyst-15, MeOH, THF (86%);(i) Cul, Zn, MVK, EVK, or t-2,4-pentadionic acid ethyl ester, EtOH:H₂O7:3 (45%); (j) Mg, EtI, Et₂O, 0° C. (73%); (j′) MeLi, Et₂O, −78° C.(85%); (k) TPAP, NMMO, molecular sieves 4A, CH₂Cl₂ (66%); (k′) (CrO₃)Py₂(“Collins”), CH₂Cl₂ (35%); (l) (1) [Ph₃PCH₂OCH₃]⁺Cl⁻, nBuLi, ether, −30°C., (2) HCl 2N, THF (48%); (m) KOH, isoprenylchloride, 18-Crown-6,toluene, ultrasound (40%); (n) KOH, allyl bromide, 18-Crown-6, THF(75%); (o) (1) Hg(OAc)₂, H₂O, THF; (2) NaBH₄, NaOH (94%); (p) SO₃.Py,Et₃N, CH₂Cl₂:DMSO 1:1 (71%); (q) (1) 9-BBN, THF, 60° C.; (2) H₂O₂, NaOH(95%); (r) (1) PDC, DMF, 40° C.; (2) CH₂N₂, Et₃O, 0° C. (36%); (s) Mg,EtI (2 eq), Et₂O, 0° C. (92%); (t) MEMCI, DIPEA, CH₂Cl₂ (80%); (u) (1)NaNO₂, DMF, urea, 25° C. (45%); (2) NaOMe (1.3 eq), MeOH; (3) O₃, Na₂S,−78° C. (70%); (v) (EtO)₂P(O)CH₂CH═CHCOOEt, LDA, THF (91%); (w) H₂/Pd (4atm), 3 h (80%); (x) Me₂BBr, ClCH₂CH₂Cl:CH₂Cl₂ 1:6 (93%); (y) Mg, MeBr,THF; (z) TBAF, THF

Scheme 6

On the other hand base induced elimination of iodide 6.2 afterprotection of the hydroxyl group gives the olefin 6.3. Hydroboration of6.3 leads to two diastereomers in a 1:1 ratio. After separation, theisomer 6.6 was transformed into the iodide 6.7. As described for theepimer 6.2, 6.7 was used to synthesize the key-intermediate 6.16.

Oxidative cleavage of the double bond in 6.3 and reduction of theresulting ketone leads to the epimeric alcohols 6.4 and 6.5. The mixtureis subjected to a Williamson ether synthesis affording the allylicethers 6.17α and 6.17β. Water-addition to the double bond, hydrolysis ofthe MOM-ether and oxidation of the resulting primary alcohol gives theepimeric aldehydes 3 6.19α and 6.19β which can be separated by HPLC(hexane-aceton 9:1). The respective structures of both epimers wereestablished by nOe measurements. 1-C homologation as already describedfor 6.10 leads to the intermediates 6.21α and 6.21β.

Also reaction of the mixture of the anions of 6.4 and 6.5 with allylbromide yields the mixture of 6.18(α+β). A sequence involvinghydroboration of the terminal double bond, oxidation and treatment withdiazomethane leads to the corresponding carboxylic methyl ester which isreacted with ethyl magnesium bromide. Subsequent hydrolysis of the MOMether and oxidation of the primary alcohol gives the epimeric aldehydeswhich are separated by HPLC. The respective structures of 6.20α and6.20β were established by nOe measurements. 1-C homologation then givesrespectively 6.22α and 6.22β. Coupling of the aldehydes 6.12, 6.13, 6.166.21α, 6.21β, 6.22α and 6.22β with the A-ring is described below.

Also transformation of iodide 6.2, via the corresponding nitro compound(39), into the aldehyde 6.24 allows introduction of the side chain. Thiscan be performed by a Horner-Wittig type reaction involving aphosphonocrotonate followed by catalytic hydrogenation. The 1-Chomologation is then carried out as described for 6.12. Coupling (24) ofthe resulting 6.26 with the anion of 13.1 leads to intermediate 6.27.Subsequently the ester function can be transformed into tertiaryalcohols. This sequence is an example of construction of analogues wherethe required side chain is formed subsequent to the Lythgoe coupling.

In another example of this series the iodo-compound 6.2 is coupled undersonication conditions with the ethyl ester of trans-2,4-pentadionicacid. Subsequent to hydrogenation of 6.28, the resulting alcohol 6.29 ishomologated to precursor 6.30 as already described.

Another example of analogues of type IIIc has an aromatic ring and caneasily be constructed from 3-hydroxyphenethyl alcohol 7.1 (scheme 7) andinvolves construction of the side chain via the phenolic hydroxyl groupand oxidation of the primary alcohol to an aldehyde function suitablefor coupling with the A ring part. Ether formation with the tosylate 7.2gives 7.3.

(a) KOH, DMSO, 4 h, r.t. (85%); (b) Et₃N, SO₃.C₅H₅N, 15 min (48%); (c)CH₃l, KO-t.Bu (55%).

Scheme 7

After oxidation of the primary alcohol in 7.3, the resulting aldehyde isbis-methylated affording the precursor 7.4.

Again several methods are possible for the synthesis of analogues withthe general structure IIIc. A few possibilities are shown in scheme 8.

In a first approach the previously described compound 3.4 (scheme 3) isetherified as before; after deprotection to the alcohol the twodiastereomers of 8.1 can be separated. Both separated alcohols 8.1α and8.1β are treated with mercuric acetate/sodium borohydride, and aresubsequently oxidized yielding the aldehydes 8.2 and 8.3, which afterthe usual coupling sequence give the analogues 22 and 23, respectively.

The β-epimer 8.1β can also be converted to a diastereomeric mixture ofepoxides which after oxidation lead to aldehyde 8.4. This is thesubstrate for coupling to analogue 24.

Finally 8.4 can also lead to en epimeric mixture of primary alcohols viaoxidation to the corresponding ketone, Wittig reaction with methylenetriphenylphosphorane and 9-BBN oxidation. After tosylation of theprimary alcohol the side chain is introduced via displacement with theanion of 3-ethoxyethyl-3-methyl-1-butyne; deprotection gives 8.5 as amixture of epimers which can now be separated. Oxidation of the α-epimer8.5α with PDC leads to aldehyde 8.6, the precursor of analogue 25.

(a) ClCH₂CH═C(CH₃)₂, NaH (89%); (b) (nBu)₄NF (81%); (c) Hg(OAc)₂; NaOH,NaBH₄ (76%) 2:1 mixt.; (d) PDC, CH₂Cl₂, r.t. (80%); (e) mCPBA, CH₂Cl₂,0° C. (86%); (f) PDC, CH₂Cl₂ (73%); (g) PDC, CH₂Cl₂ (96%); (h)Ph₃P⁺CH₃Br⁻, nBuLi, THF (83%); (i) 9-BBN (90%); (j) TsCl, pyridine(95%); (k) HC≡CC(Me)₂OEE, NaH, DMSO (62%); (l) (nBu)₄NF, THF (92%); (m)PDC, CH₂Cl₂ (71%).

(aa) t.butyldimethylsilyl ethyl ketene acetal, Hgl₂, CH₂Cl₂; (ab)LiAlH₄, Et₂O; (ac) TBAF, THF (61% from 8.1); (ad) TBDMSCl, imidazole,DMF (99%); (ae) O₃, MeOH, −30° C., FeSO₄, Cu(OAc)₂; (af) Pd, H₂ (4 atm)(61% from 8.2); (ag) TBAF, THF (100%); (ah) MEMCl, EtiPr₂N, CH₂Cl₂(99%); (ai) NaBH₄, MeOH (70%); (aj) KOH, 18-crown-6,chloro-3-methyl-2-butene, toluene, ultrasound (43%); (ak) Hg(OAc)₂/NaOH,NaBH₄ (78%); (al) Amberlyst-15, MeOH:THF 1:1 (100%); (am) CH₂Cl₂:DMSO1:2, pyridinesulfurtrioxide complex, Et₃N (69%).

(ba) K₂CO₃, MeOH, 1 h, r.t. (55%); (bb) BnO—C(═NH)CCl₃, CF₃SO₃H,CH₂Cl₂/c.hexane, 90 min, 0° C. (60%); (bc) (i) FOSMIC, BuLi, Et₂O, 2hrs, 0° C.; (ii) HCl (37% soln), 12 hr, r.t. (67%); (bd)Ø₃P═CH—CH₂—COO—, THF, 2 h, r.t.; (be) CH₂N₂, Et₂O (28% overall); (bf)MeLi, LiBr, diethyl ether, 2 hr, 0° C.; (bg) Pd/C 10%, EtOAc, H₂, 6 hr,r.t. (53%); (bh) NMMO, TPAP, CH₂Cl₂, 2 h, r.t. (85%).

Scheme 8

An example of the synthesis of analogues of general formula IIIcstarting from R-carvone (8.7) is also shown in scheme 8. The strategycenters around (a) diastereoselective 1,4-addition (b) removal of theisopropylidene group (40) (c) introduction of an oxa-side chain. Thisroute leads to separable diastereoisomers.

The 1,4-addition involving a silylated ketene acetal on 8.7 leads to anenol silyl ether. The ester function in this intermediate isconveniently reduced to a hydroxyl function prior to hydrolysis.Ozonolysis of 8.8 and subsequent treatment with iron and copper saltsallows cleavage of the isopropylidene substituent. Catalytichydrogenation of the resulting double bond and changing the protectivegroup gives the MEM ether 8.10. Subsequently sodium borohydridereduction leads to isomeric alcohols 8.11. This mixture is subjected toether formation with isoprenylchloride. The ethers 8.12, 8.13 and 8.14can be separated. Each is individually transformed in the tertiaryalcohols 8.16, 8.17 and 8.18 respectively.

Still another method for obtaining analogues of general formula IIIc canbe illustrated starting from compound 8.19, a ketone described in theliterature (41). It involves side chain construction making use of thecarbonyl function.

Reaction with diethyl(isocyanomethyl)phosphonate followed by acidhydrolysis gives the aldehyde 8.21. Wittig homologation introduces theside chain. Reaction of methyllithium on 8.22 leads to the tertiaryalcohol. The double bond is hydrogenated with concomitant cleavage ofthe benzyl ether. Finally oxidation of the primary hydroxyl group in8.23 gives the precursor aldehyde 8.24.

An example of the synthesis of an analogue of general formula IIIe isshown in scheme 9. Starting from the known homochiral enone 9.1 (42) adissolving metal ammonia reduction leads to the trans-fused decalone9.2. The introduction of the side chain involves reaction with thesodium salt of protected 2-methyl-3-butyn-2-ol, followed by dehydrationto 9.3. Catalytic hydrogenation eventually leads to decalone 9.4, theprecursor of analogue 31.

(a) Li, I.NH₃, (56%); (b) NaC≡C—C(Me)₂OEE, DMSO (74%); (c) Tf₂O, CH₂Cl₂,py, DMAP (25%); (d) H₂, Pd, EtOAc (65%); (e) TMS, imidazole;

(aa) Hgl₂, CH₂(OTBAS)(OEt), Et₃N, CH₂Cl₂, 3 h, −78° C.—r.t. (97%); (ab)toluene, glycol, H₂SO₄, molecular sieves 3 Å, 10 h, reflux (75%); (ac)DIBAH, toluene, 4 h, −78° C. (93%); (ad) triethylphosphonoacetate, BuLi,THF, 17 h, −78° C.—r.t. (88%); (ae) 10% Pd/C, hexane, 1 atm H₂, 1.5 h,0° C. (99%); (af) MeMgl, diethylether, 5 h, r.t. (85%); (ag)Amberlyst-15, THF:water 2:1, 12 h, r.t. (99%); (ah) TSIM, THF, 2 h, r.t.(97%); (ai) EtMgl, diethylether, 2 h, r.t. (89%); (aj)Ph₃P⁺(CH₂)₃COOBzBr—, LDA, HMPA:THF 1:1, 2 h, −20° C. (21%);

Scheme 9

Further examples of analogues of general formula IIIe whereby one of therings of the bicyclic system is a heterocyclic ring are also shown inscheme 9. The synthesis starts from the known enone 9.5 (28) andproceeds via conjugate addition, heterocyclic ring formation and Wittigcondensation as shown in the scheme. Various carbonyl derivatives wereobtained that were condensed with the A-ring in the usual way.

Examples of precursors for analogues of type IIIb, with a cyclohexanoicD-ring, are described in scheme 10. The starting material for theseparticular examples is the known 10.1 (43); the ester function is thehandle for the side chain construction while the carbonyl function canbe transformed into a formyl group. Alkylation of 10.2 leads to 10.3 asthe major (95%) epimer in accordance with literature precedents (44).After transformation of the ester function to a methyl group, followinga classical procedure, the terminal double bond in 10.6 is cleaved byozonolysis. Finally deprotection leads to ketone 10.7.

(a) PPTS, acetone, 2 h, reflux (86%); (b) LDA, THF, 1 h, −30° C.;5-Br-1-pentene, HMPA, 3 h, −78° C. (93%); (c) LiAlH₄, Et₂O (99.8%); (d)TosCl, TEA, DMAP, DCM, 20 h, r.t. (95%); (e) LiAlH₄, Et₂O, 5 h, reflux(88%); (f) O₃, DCM:2.5M NaOH in MeOH 4:1 (v/v), 45 min, −78° C. (64%);(g) PPTS, acetone, H₂O (cat), 3 h, reflux (75%); (h) FOSMIC, BuLi, Et₂O,15 min, −60° C.; HCl 37%, 12 h, r.t. (64%); (i) Me₂S═CH₂, THF, 2 h, r.t.(33%); (j) BF₃.OEt₂, Et₂O, 12 h, r.t. (65%).

Scheme 10

The formation of a formyl substituent from a ketone is well known. Twomethods are used here; one of which involving reaction with diethyl(isocyanomethyl)phosphonate (45). The epimeric aldehydes 10.8 and 10.9can be separated. Also base catalyzed epimerization of 10.9 gives thethermodynamically more stable 10.8. Both precursors 10.8 and 10.9 can betransformed into analogues via coupling with 13.1 and organometallicreactions under conditions similar to the synthesis of 19 from 6.27. Theother method involves the intermediacy of the epoxide 10.10 which isthen transformed into the mixture of 10.8 and 10.9.

Examples of precursors for compounds of type IIb with a 5-memberedD-ring are described in scheme 11.

In one case the synthesis starts from the t-butyldimethylsilyl ether11.1 of the commercially available 5-(hydroxymethyl)furfural. Wittigreaction with the ylid 11.2 yields the ester 11.3 which is easilytransformed into the tertiary alcohol 11.4. Finally deprotection andoxidation of the primary hydroxyl group affords the precursor 11.5.

The precursor 11.11 can be obtained from the known 11.6 (46) andinvolves hydroboration of the double bond after reductive removal of thebromo-atom and formation of the tosylate. The epimers 11.9 are thencoupled with the side chain. Oxidation yields the epimeric aldehydes11.11α+11.11β.

A closely related precursor can be obtained from (−)-camphoric acid(11.12). Subsequent to reduction, SAM II lipase catalyzed mono-esterformation allows for the requisite differentiation of the two hydroxylfunctions. From 11.13, subsequent to oxidation to the correspondingaldehyde, the side chain can be introduced. This leads to intermediate11.14.

On the one hand, Grignard reaction and oxidation of the primary alcoholleads to precursor 11.21. On the other hand 11.14 can easily betransformed into precursors 11.19 and 11.20; now an additional catalytichydrogenation step is involved.

Another D-ring analogue of type IIb, namely 8,9-seco-1α,25-(OH)₂ vitaminD₃ is available from 11.22 (from 12.1). Formation of an enol derivative(e.g. the triflate) via the kinetically produced enolate anion andsubsequent ozonolysis gives 11.24. Reduction of the correspondingtosylate 11.25 and subsequent oxidation of the primary hydroxyl group in11.26 affords the 8,9-seco C/D ring precursor 11.27.

(a) THF, HMPA, 2 h, −20° C. (62%); (b) EtMgBr, Et2O, 5 h, −10° C. (86%for 11.14; 75% for 11.17); (c) TBAF, THF, 1 h, r.t.; (d) SO₃-pyridine,CH₂Cl₂, DMSO, 3 h, −10° C. (40% from 11.4; 63% from 11.10; 80% from11.13); (e) nBu₃SnH, 100° C.; (f) TsCl, Et₃N, CH₂Cl₂, DMAP 71%; (g) (i)9-BBN, THF, 60° C.; (ii) H₂O₂, NaOH (85%); (h) ≡—C(Et)₂OEE, NaH, DMSO,90 min, 65° C. (63%); (i) LiAlH₄, THF, Et₂O, 4 h (88%); (j) vinylacetate, SAM II, 66 h, 37° C. (60%); (k) tri-ethyl-4-phosphonoacetate,LDA, THF, 24 h. 0°→25° C.; (l) K₂CO₃, EtOH, r.t. (65% overall); (m) 5%Rh/Al₂O₃, EtOAc, H₂ (90%); (n) MeMgBr, Et₂O, 90 min, r.t. (86% for11.18, 94% for 11.15); (o) TPAP, NMNO, CH₂Cl₂, 2 h, r.t. (80-78%); (p)LDA, THF, 15 min, −78° C., 2 h, r.t., then PhNTf₂, 18 h, 0° C. (65%);(q)O₃, NaHCO₃, MeOH, −78° C., then NaBH₄, MeOH, 18 h, −78° C. to r.t. (91%overall); (r) LiAlH₄, THF, Δ, 36 h (61%); (s) TPAP, NMNO, CH₂Cl₂ ₁, h,r.t. (50%);

Scheme 11

Examples of precursors for the synthesis of analogues of type IId andwhich are characterised by a cis-fused bicyclic system are shown inscheme 12. These precursors can be obtained via (a) ozonolysis ofvitamin D₂, (b) introduction of a side chain and (c) epimerisation atC-13. Epimerisation of the known ketone 12.1 (47) leads to a circa 3:1ratio in favour of the cis-fused isomer. The 25-hydroxyl group isprotected prior to coupling with the A-ring. It is also possible tostart from the known inhoffen-Lythgoe diol (48) which can easily betransformed into the monotosylate 12.3. Reaction of 12.3 with the anionof 3-ethoxyethyl-3-methyl-1-butyn leads to 12.4 an intermediate for twoprecursors. Oxidation and epimerisation afford the ketone 12.5.

On the other hand elimination of the 25-oxy-function leads to 12.6 inwhich the double bond can selectively be epoxidised. Oxidation of thehydroxyl group and subsequent DBU mediated epimerisation gives cis-fusedketone 12.7.

(a) NaOMe, MeOH, 24 hrs, r.t. (73% for 12.2; 65% for 12.5); (b) TMSimidazol, CH₂Cl₂, 3 hrs, r.t. (79%); (c) NaH, DMSO, HC≡C—C.(CH₃)₂OEE(67% for 12.4; 56% for 12.12); (d) PDC, CH₂Cl₂, 2 hrs (84% for 12.5; 69%for 12.7; 70% for 12.12); (e) TsOH, toluene, 60° C. (74%); (f) mCPBA,Na₂HPO₄, THF (81%); (g) DBU, CH₂Cl₂, 3 d, r.t.; (h) (i) O₃, CH₂Cl₂:MeOH(1:1), −78° C.; (ii) Me₂S, r.t.; (i) 5% HCl, THF (1:3), 30° C., 36 hrs;(j) NaBH₄, MeOH, r.t. (99%); (k) TsCl, py, 0° C., 12 hrs (56%); (l)triethyl-4-phosphonocrotonate, DLA, THF, −78° C.→r.t., 3 h (85%); (m)NaOEt, EtOH, r.t., 21 h (62%); (n) H₂, Rh/Al₂O₃, EtOAc, r.t., 1.5 h(89-93%).

Scheme 12

Of importance is the fact that efficient epimerisation at C-20 and C-13can be effected simultaneously. Ozonolysis of vitamin D₂ with nonreductive work-up gives the keto-aldehyde 12.8 which upon acid catalysedepimerisation leads to a mixture of the four possible isomers from whichthe major component 12.9 can be isolated by HPLC. It is more facile toisolate the two cis fused isomers together and to reduce the carbonylfunctions before separation of the C-20 epimers. The primary hydroxylgroup in 12.10 can be tosylated with a sufficient selectivity. Couplingof the tosylate 12.11 with the anion of 3-ethoxyethyl-3-methyl-1-butynand subsequent oxidation affords the precursor 12.12. An analogouscoupling leads to precursor 12.13. These ketones and thetetrahydroderivative 12.14 can be coupled with the anion of 13.2affording respectively analogues 46, 48 and 47.

Selective Horner-Wittig reaction of the aldehyde function in 12.9 withthe anion of triethyl 4-phosphonocrotonate is an alternative for sidechain construction. This leads to 12.15 and subsequently to 12.16.Coupling with 13.2 followed by reaction with an appropriate organometalleads to analogues 49 to 52. The same sequence, but starting from theS-epimer 12.8 leads to 12.17 and 12.18 precursors for analogues 53 to55.

The precursor aldehydes or ketones described in schemes 1, 2, 3, 5, 6,7, 8, 9, 10, 11 and 12 are coupled with the A-ring phosphine oxides 13.1and 13.2 using the Lythgoe procedure (scheme 13). In this manner thevitamin D₃ analogues 1 to 55 shown in table I are obtained. With respectto the 5- and 6-membered rings of type C, D and E, and combinations CD,CE and DE (see table 1), it is noted that the rings may be saturated,such as cyclopentane or cyclohexane, unsaturated such as cyclopentene orcyclohexene.

Scheme 13

R—CHO + 13.1 a.c 1 1.8d + 13.2 a.c 2 1.11d + 13.1 a.b 3 2.5 + 13.1 a.b 42.7 + 13.1 a.b 5 2.7 + 13.2 a.b 6 2.12 + 13.1 a.b 7 2.12 + 13.2 a.b 83.7 + 13.1 a.b 9 5.6 + 13.1 a.b 10 6.12 + 13.1 a.b 11 6.13 + 13.1 a.c 126.13 + 13.2 a.c 13 6.21α + 13.1 a.c 14 6.22α + 13.1 a.c 15 6.16 + 13.1a.c 16 6.21β + 13.1 a.b 17 6.22β + 13.1 a.c 18 6.26 + 13.1 a.e.b 196.26 + 13.1 a.f.b 20 6.3 + 13.1 a.e.b 21 8.2 + 13.1 a.b 22 8.3 + 13.1a.b 23 8.4 + 13.1 a.b 24 8.6 + 13.1 a.d.b 25 8.18 + 13.1 a.b 26 8.16 +13.1 a.b 27 8.17 + 13.1 a.b 28 8.24 + 13.1 a.b 29 7.4 + 13.1 a.d.b 309.4 + 13.2 a.b 31 9.9 + 13.1 a.b 32 9.11 + 13.1 a.b 33 9.12 + 13.2 a.b34 9.14 + 13.2 a.b 35 9.13 + 13.1 a.b 36 11.19 + 13.1 a.b 37 11.20 +13.1 a.b 38 11.21 + 13.1 a.b 39 11.27 + 13.1 a.b 40 11.11 + 13.1 a.c 4111.5 + 13.1 a.b 42 12.2a + 13.2 a.b 43 12.5 + 13.2 a.c 44 12.7 + 13.2a.c 45 12.12 + 13.2 a.c 46 12.14 + 13.2 a.c 47 12.13 + 13.2 a.c 4812.15 + 13.2 a.e.c 49 12.15 + 13.2 a.f.c 50 12.16 + 13.2 a.e.c 5112.16 + 13.2 a.f.c 52 12.17 + 13.2 a.e.c 53 12.18 + 13.2 a.e.c 5412.18 + 13.2 a.f.c 55 10.8 + 13.1 a.e.b 56 10.9 + 13.1 a.e.b 57 (a)n.BuLi, THF −78° C.; (b) n.Bu₄NF, THF; (c) Amberlyst-15, MeOH; (d) PPTS,CH₂Cl₂; (e) MeMgX, THF, r.t.; (f) EtMgX, THF, r.t..

The rings may also be substituted with one or more substituents selectedfrom the group comprising alkyl, alkenyl, alkynyl, aryl, halogen,hydroxy and functional groups derived therefrom such as ethers andesters, and amine and functional groups therefrom such as N-alkylatedamines and amides.

The Horner-Wittig coupling using the classical A-ring phosphinoxide andthe trans-fused CD-ring ketone leads exclusively to theE-stereochemistry at the 7,8-double bond ( ). The profound modificationof the central CD-ring system in the new analogues described above canresult in a change in stereoselectivity for that transformation. This isespecialy true in cases where the Wittig condensation is performed oncycloalkanones of which the α-positions may be less differentiatedcompared to the classical example. Hence this problem may be expectedespecially in the case of the synthesis of analogues of type IIIa, IIIdand IIIe. As an example the Wittig condensation on decalone 9.4 leads toa 2:1 mixture of E- and Z-derivatives 14.1 and 14.2 that are furtherhydrolyzed to analogue 31 that is isolated as a mixture of 2:1 isomers.A similar example is the reaction on 3.7 which led to a separable 4:1mixture of E:Z-isomers 14.3 and 14.4.

Also in other cases, however, can this stereoselectivity problem occur.As an example the Wittig condensation on aldehyde 11.27 leads to a E:Zmixture of 14.5 and 14.6 that can be separated, one of which leadingafter hydrolysis to analogue 40.

At higher temperatures vitamin D derivatives possessing the naturaltriene system are known to rearrange readily into the so-calledprevitamin D derivatives (scheme 15). In the natural series the vitaminD structure predominates in the equilibrium (approximate ratio at 25°C.=9:1). A substantial change in the CD-ring part of the molecule may,however, affect considerably this equilibrium composition. Also, theconversion of the vitamin form into the previtamin form may occur morereadily than in the natural derivatives.

As an example the carbonyl derivative 2.7 was found to lead, after theusual Wittig-Horner coupling and a somewhat difficult hydrolysis ofsilylprotective groups (40° C., 40 h; TBAF in THF) to a mixture ofanalogue 5 and its corresponding previtamin form, compound 58.

For certain types of analogues the presence of a 19-nor A-ring ismandatory. Ketones, of type VII, when used as precursors of 19-noranalogues can be coupled, using the Lythgoe procedure, with 13.2, anexample of phosphine oxide IV, or alternatively with alkynes of type VI.It is also possible to transform ketones VII into vinylic bromide VIIwhich can react with the carbonyl function in V. The 19-Nor-A ringprecursors V and VI are alternatives for 13.2 can be obtained from(−)-quinic acid 16.1. The method is based on the “cyclovitamin” strategyfor which there are examples in the case of the natural series (19methylene). The two essential features are the simultaneous removal ofthe 1- and 4-hydroxyl functions in 16.1 and formation of thebicyclo[3.1.0]hexane skeleton. The 5-hydroxyl group in lactone 16.2 isprotected, for instance as a t-butyldimethyl silyl ether; 16.3 can beseparated from the minor regioisomer. The two hydroxyl groups areremoved by the Barton-McCombie deoxygenation via the bis-thiocarbonylimidazolide 16.4, as one of the several potential methods (29).Solvolysis of the resulting 16.5 gave 16.6. Transformation of thehydroxyl function into a suitable leaving group and subsequentbase-induced cyclopropane formation gave ester 16.8. The two precursors16.10 and 16.11 are now readily available; one of the possible methodsfor alkyne formation is reaction of aldehyde 16.10 with dimethyldiazomethylphosphonate (35). Coupling of 16.11 with an appropriateketone of type VII (such as 12.2b) can be carried out as described inthe natural series and comprises reaction of the anion of 16.11, LiAlH₄reduction of the resulting propargylic alcohol unit and acid catalyzedsolvolysis giving the 19-nor vitamine analogue 43.

Aldehyde 16.10 can also directly be used via reaction of a appropriatevinylic anion derived from a vinylic halide of type VII (such as 16.12).The vinylic halide is accessible from a ketone with for instance aWittig type olefination.

(a) TsOH, toluene, Δ, 15 h (79%); (b) TBDMSCl, imid, DMAP, DMF, r.t., 12h (66%); (c) (imid)₂C═S, DMAP, Δ, 3 d (87%); (d) Bu3SnH, AIBN, toluene,Δ, 5 h (55%); (e) NaOMe, MeOH, 0° C., 1 h (100%); (f) p-BrC₆H₄SO₂Cl,CHCl₃, py, 0°—r.t., 13.5 h (100%); (g) t-BuOK, t-BuOH, Δ, 1 h (71%); (h)DIBAH, toluene, −78° C., 2 h (98%); (i) PCC, CH₂Cl₂, r.t., 2 h (90%);(j) (MeO)₂P(O)CHN₂, t-BuOK, −78° C.→r.t., 18 h (89%); (k) 16.12, t-BuLi;Et₂O; −78° C.; 50 min; 16.10, 1 h (46%); (l) p-TsOH, H₂O-dioxane (1:3),63° C., 6 h (78%); (m) Ph₃P⁺CH₂Br; Br⁻; NaN(TMS)₂, THF, −68° C., 1 h;12.2, −68° C., 1 h, r.t. overnight (56%); (n) 16.11, n-BuLi, THF, −50°C., 1 h, 12.2, r.t., 30 min (55%); (o) LiAlH4, NaOMe, THF, reflux, 2 h,(50%); (p) p-TsOH; H₂O-dioxane (1:3), 63° C.; 6 h (40%).

Scheme 16

!

Several analogues of vitamin D related to this invention, arecharacterized by a central part whose structure has been thoroughlymodified, and yet maintain a biological activity similar to vitamin D.Especially those derivatives which lack the combined presence of thesix- and of the five-membered ring typical of the vitamin D skeleton,and which can be considered as non steroidal analogues of vitamin Dconstitute the first examples of an entirely novel series of vitamin Danalogues.

In particular it appears that the classical trans-fused perhydrindaneCD-ring system is not in se necessary for biological activity. In thisrespect it was also discovered that steroidal analogues possessing theunnatural cis-fused CD-ring system were in fact active; in these cases,however, the structure of the A-ring should not allow for possiblepreferential rearrangement to the previtamin D form.

Finally, it also appears that the presence of certain conformationallyrestricting structural features, such as rings and/or alkyl substituentswithin the central part are necessary, since the derivative (1) with alinear unsubstituted central chain is not active.

We found that the compounds described above and belonging to a new classof drugs, including vitamin D analogues with modifications of the CDring structure, have a selective activity on cell function, such asinhibition of cell proliferation (non-malignant cells such askeratinocytes as well as malignant cell such as breast carcinoma,osteo-sarcoma and leukemia cells) and also have a high potency forinduction of cell differentiation (e.g. cell types as just mentioned)but on the other hand have strikingly lower effect on calcium and bonehomeostasis as evaluated in rachitic chicks (by measuring serum and bonecalcium, and by measurement of two vitamin D-dependent proteins, serumosteocalcin and duodenal calbindin D) as well as in vitamin D repletednormal mice (using similar end points). Thus, unlike the classicalvitamin D compounds, the new drugs do not have the same toxic effect oncalcium and bone homeostasis. In light of prior art and studies it wasunexpected and surprising that the central part of the classical vitaminD structure, known as the CD ring, is not essential for all actions ofthe vitamin D hormone and that on the contrary modifications in thispart express selective activities of the spectrum of vitamin D activitythat can be used therapeutically for several disorders. Specifically thenew drugs can be used for the therapy or prevention of

immune disorders, such as autoimmune diseases (such as, but not limitedto diabetes mellitus type 1, multiple sclerosis, lupus and lupus likedisorders, asthma, glomerulonephritis, etc.) selective dysfunctions ofthe immune system (e.g. AIDS) and prevention of immune rejection [suchas rejections of grafts (e.g. kidney, heart, bone marrow, liver, isletsor whole pancreas, skin etc.) or prevention of graft versus hostdisease]. The newly invented drugs can either be used alone or incombination with other drugs known to interfere with the immune system(e.g. cyclosporin, FK 506, glucocorticoids, monoclonal antibodies,cytohines or growth factors . . . ). In analogy with the immune activityof the new compounds, similar effects can be expected in otherinflammatory diseases (e.g. rheumatoid arthritis).

skin disorders either characterized by hyperproliferation and/orinflammation and/or (auto)immune reaction (e.g. psoriasis, dyskeratosis,acne). Moreover since these drugs can stimulate the differentiation ofskin cells they can be used for the treatment or prevention of alopeciaof different origin including alopecia due to chemotherapy orirradiation.

hyperproliferative disorders and cancer such as hyperproliferative skindiseases (e.g. psoriasis) and several types of cancers and theirmetastases (all types of cancer which have or can be induced to havevitamin D receptors such as but not limited to breast cancer, leukemia,myelo-dysplastic syndromes and lymphomas, squamous cell carcinomas andgastrointestinal cancers, melanomas, osteosarcoma . . . ). The newlyinvented drugs can, again as for the other indications, be used alone inthe appropriate form and route of administration or used in combinationwith other drugs known to be of therapeutic value in such disorders.These new drugs may be particularly advantageous for such diseases asthey can, in contrast to classical chemotherapeutic agents, alsostimulate cell differentiation.

endocrine disorders since vitamin D analogues can modulate hormonesecretion, such as increased insulin secretion or selective suppressionof parathyroid hormone secretion (e.g. in chronic renal failure andsecondary hyperparathyroidism).

diseases characterized by abnormal intracellular calcium handling sincethe new drugs have favourable effects in cells whose functions dependlargely on intracellular calcium movements (e.g. endocrine cells, muscle. . . ).

The use of the new compounds can find application as well in humandisorders as in veterinary medicine.

The amount of the new compounds necessary for their therapeutic effectcan vary according to its indication, route of administration andspecies (animal/man) treated. The compounds can be administered byenteral, parenteral or local topical route. In the treatment ofdermatological disorders a topical application as ointment, cream orlotion is to be preferred over systemic treatment, preferably in a doseof 0.1 to 500 μg/g. The systemic administration as tablets, capsules,liquid or as sterile preparation in an appropriate carrier, diluentand/or solvent for parenteral injection will use microgram quantities ofthe compounds per day depending on the indication and theclinical/veterinary situation.

The advantage of the new compounds over the natural or existing vitaminD metabolites or analogues is due to their intrinsic activity ininduction of cell differentiation, inhibition of cell proliferation andmodulation of the cellular activity in general, while neverthelessdisplaying reduced calcemic effects in vivo. Indeed such calcemiceffects, present in other vitamin D metabolites or analogues are to beconsidered as undesired side effects since the doses required for toabove mentioned indications are sometimes supraphysiologic and wouldresult in serious calcemic abnormalities when other vitamin Dmetabolites or analogues would be used.

Biological Evaluation of the Novel Vitamin D Analogues

1. Binding Properties of the New Novel Vitamin D Analogues

The methods used to evaluate the binding properties of the new analoguesare examples of the state of the art techniques used for steroid hormone(including vitamin D) binding assays as described previously.

The affinity of the analogues of 1α,25-(OH)₂D₃ to the vitamin D receptorwas evaluated by their ability to compete with [³H]1α,25-(OH)₂D₃(specific activity 180 Ci/mmol Amersham, Buckinghamshire, UK) forbinding to the high speed supernatant from intestinal mucosa homogenatesobtained from normal pigs (22,23). The incubation was performed at 4° C.for 20 h and phase separation was obtained by addition of dextranoatedcharcoal. The affinity for 1α,25-(OH)₂D₃ was 1.06±0.38×10¹⁰ M⁻¹ (M±SD,n=10). The relative affinity of the analogues was calculated from theirconcentration needed to displace 50% of [³H]1α,25-(OH)₂D₃ from itsreceptor compared with 1α,25-(OH)₂D₃ (assigned a 100% value). (Table 2).

The relative affinity for hDBP was measured by incubating[³H]1α,25-(OH)₂D₃ and increasing concentrations of 1α,25-(OH)₂D₃ or itsanologues with purified hDBP (0.2 μM) in 1 ml (0.01 M Tris-HCl, 0.154 MNaCl, pH 7.4) for 3 h at 4° C., followed by phase separation by additionof cold dextran-coated charcoal (2,23).

The results obtained with some examples of the new analogues are givenin Table 2. These data clearly show a binding to the vitamin D receptor,necessary for their biological activity, while their binding for thevitamin D binding protein, known as DBP, is decreased in comparison with1α,25-(OH)₂D₃. We and others have previously demonstrated for othervitamin D analogues that such reduced binding to DBP enhances its ratioof cell differentiating over calcemic effects (23,37).

2. Effects of the Novel Vitamin D Analogues on Cell Proliferation andCell Differentiation

The cell culture systems were used according to the state of the art

to evaluate the effects on cell proliferation of non-malignant cells andespecially to evaluate their potential for use for dermatologicaldisorders, the new compounds were tested in cultures of human normalkeratinocytes.

Human skin keratinocytes were isolated and cultured using a modificationof the method of Kitano and Okada (38).

Briefly, the skin from biopsies of patients with breast tumors, wascutted into pieces measuring 3-5 mm and soaked overnight at 4° C. in asolution of dispase (20 Boehringer units/ml). The epidermis was peeledfrom the dermis, washed with calcium- and magnesium-free phosphatebuffered saline and incubated and shaked in a 0.25% trypsin solution for10 min at room temperature. The reaction was then stopped by addition ofPBS containing 10% FCS. The cells were collected after centrifugation at4° C. for 10 min at 800 rpm. After an additional washing with PBS, thepellet was suspended in culture medium into 25 cm² primaria flasks fromBecton Dickinson. The keratinocytes were cultivated at 37° C. in anatmosphere of 5% CO₂ in air. A few hours later, the medium was replacedby new one. The medium [Keratinocyte Medium from Gibco containingEpidermal Growth Factor (5 ng/ml), Bovine Pituitary Extract (35-50μg/ml) and antibiotics] was renewed every other day until confluency.

Keratinocytes were cultured in 96-well plate and, after 24 hours, weretreated with various concentrations of vitamin D analogues, followed bypulse labelling with 1 μCi of [³H]thymidine for 3 hours. Cultures werewashed 3 times with PBS and twice with 10% (v/v) ice coldtrichloroacetic acid. Cells were solubilized with 1 M NaOH andradioactivity was counted in a scintillation counter.

to evaluate the effect on cell proliferation and induction of celldifferentiation, malignant cells were grown in vitro and theirproliferation was evaluated by measuring cell number, protein contentand the incorporation of radioactive thymidine. As examples of malignantcell human leukemia cells (HL 60), human osteosarcoma cells (MG 63cells) and both murine and human breast cancer cells (MCF 7, MFM223 andGR cells) were used. In addition the effect of the new drugs showedadditive effects when tested in combination with other anticancer drugs(e.g. retinoic acids, anti-estrogens . . . ).

HL-60 cells were seeded at 1.2×10⁵ cells/ml and 1α,25-(OH)₂D₃ or itsanologs were added in ethanol (final concentration <0.2%) in RPMI 1640medium supplemented with 10% heat-inactivated fetal calf serum (FCS) for4 d at 37° C. Cells were then assayed for maturation by NBT reductionassay as described (22) using a hemacytometer, or for proliferation bycell counting and [³H] thymidine incorporation. MG 63 cells, seeded at5×10³ cells/ml in 96 well flat bottomed culture plates (Falcon, BectonDickinson, N.J.) in a volume of 0.2 ml of DMEM and 2% FCS, wereincubated with 1α,25-(OH)₂D₃ or its analogues for 72 h. Osteocalcin wasthen measured in the culture medium using a homologous human osteocalcinRIA (39). Breast carcinoma cells (MCF-7 or GR) were grown inDMEM/nut.mix F-12 (HAM) medium supplemented with 10% FCS. Cells(5000/Well) were incubated during 24 h in 96 well tissue culture plates(Falcon 3072) followed by a 72 h incubation with/without 1α,25-(OH)₂D₃or analogues. The cells were then incubated with [³H]thymidine (1μCi/well) for 4 h and harvested thereafter in NaOH (0.1 M) and theradioactivity counted. The protein content of the cells was measured bythe Pierce BCA protein assay (Rockford, Ill.).

Results obtained with some of the new analogues are presented in Table 2and FIGS. 1-5.

to evaluate the immune potential of the new drugs their biologicalactivity was tested in a mixed lymphocyte test in vitro according tostate of the art procedures; in addition the effects of the analoguesfor induction of differentiation of HL 60 cells into mature monocyteswas tested in vitro. Moreover their immune potential was demonstrated invivo by their potency to decrease the graft versus host reaction in miceand to prevent the neurological events in a mice model of experimentalallergic encephalitis.

The capability of the novel analogues to activate the genomic pathwaynormally used by natural vitamin D metabolites was demonstrated bytransfection studies using a construct of several direct repeats ofvitamin D responsive elements (using the mouse osteopontin or ratosteocalcin VDRE sequence coupled to a CAT or hGH reporter gene(constructs made by J. White and G. N. Hendy, Montreal, Canada and M. R.Haussler, Tucson, Ariz.).

TABLE 2 Summary of the biological properties of some selected newvitamin D analogues: affinity for the vitamin D receptor/plasma vitaminD-binding protein, their potency to induce celldifferentiation/inhibition of cell proliferation in human leukemia(HL-60), osteosarcoma (MG-63), human breast cancer (MCF-7) cells andhuman keratinocytes. Compound Affinity for Cell differentation/cellproliferation Ca serum number pig receptor human DBP HL-60 MG-63 MCF-7keratinocytes chick  1 0.1 0.1 0.1 0.2 / / /  2 0 0 75 4.5 / / /  3 10010 720 75 / / /  4 60 5 500 100 2000 2000 <1  5 80 10 4000 1000 60005000 9  6 35 2 3000 1000 1000 / 18  7 60 20 1000 700 6000 / 5  8 453 >1000 300 3000 / <0.1 11 8 19 30 35 30 10 <0.1 12 45 5 250 135 80 100<0.1 16 15 0 12 10 / 30 0.2 18 4 0 10 / 40 35 <0.1 19 13 50 10 30 / / /20 28 8 75 30 / / / 38 80 2 100 / / / 2 43 25 7 40 100 20 100 0.1 44 262 320 150 1000 700 0.1 45 2.5 1.5 80 10 20 / 1 46 30 <0.5 1650 500 19501000 2 47 80 <0.5 500 450 1050 1000 10 48 40 0.3 2200 800 1500 350 1 509 0 40 / 100 60 0.1 51 100 2 600 400 600 400 0.8 52 100 0 600 900 2000300 1.6 54 20 1 30 10 / / 0.4 55 30 0 65 3 / / 0.3 56 / 70 100 / / / /58 3 1 650 115 700 1000 <1 Biological values are expressed as % of theactivity of 1α,25-(OH)₂D₃ at 50% of its activity (B50). For detail ofthe methods see text. The numbering of the compounds is identical to thenumbers used for the description of their chemical structure in Table 1.

3. In vivo Evaluation of the Immune Potential

To evaluate the immune potential of the analogues the well known modelof prevention of autoimmune disease recurrence in the spontaneouslydiabetic NOD mice was used. When syngeneic NOD islets are transplantedunder the kidney capsule of spontaneously diabetic NOD mice, diabetes isonly cured for some days, since in the absence of immunomodulatorytreatment, the newly transplanted islets are destroyed within 14 days.Cyclosporin A, a well known immunosuppressant, can only delay recurrenceat near toxic doses (15 mg/kg/d). Combination of subtherapeutical dosesof CyA (7.5 mg/kg/d) with low, noncalcemic dosis of one of the newanalogues (number 46, from table 1) (10 μg/kg/2d) a spectacularprolongation of graft survival is observed, with survival of the grafteven after discontinuation of therapy (day 60). (Table 3)

TABLE 3 Mean survival of islets and Calcemia range (d) (mg/dl) Control 8(5-13) 9.7 CyA 7.5 mg/kg/d 15 (4-42) 10.1 CyA 15 mg/kg/d >58 (22 −> 90)10.1 Nr 46 10 μg/kg/2d 19 (6-51) 8.5 Nr 46 10 μg/kg/2d + >69 (23 −> 90)9.1 CyA 7.5 mg/kg/d

4. Calcemic Effects of Novel Vitamin D Analogues

To evaluate calcemic effects in vivo tests were performed using chicksand mice.

The antirachitic activity of the analogues was tested in 3 weeks oldvitamin D-deficient chicks injected for 10 consecutive days with1α,25-(OH)₂D₃ or its analogues (22,23). Serum calcium (by atomicabsorptiometry) and osteocalcin (by specific RIA), duodenal calbindinD-28K (by RIA) and bone calcium content were measured. The hypercalcemiceffect of the most interesting anologues was also tested in vitaminD-replete normal NMRI mice by daily sc injection of 1α,25-(OH)₂D₃, itsanalogues or the solvent for 7 consecutive days, using serum, bone andurinary calcium excretion and serum osteocalcin (by specific mouse RIA)as parameters (40).

The representative data obtained with some of the new analogues arepresented in FIG. 6.

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EXAMPLE 1

Synthesis of the cis-decalone 1.2b

To a suspension of AlCl₃ (2 g, 14.99 mmol) in toluene (250 ml) at −78°C., is added 1.1. The solution was stirred for 1 h under Ar, while thetemperature raised to r.t. At a ratio of 2 ml/15 min, isoprene (11 ml;0.11 mol) in toluene (80 ml) is added with a motor driven syringe. After6 h the mixture is poured into an ice cooled saturated NaHCO₃ solution.The solution is exracted with Et₂O and the combined organic layers aredried (NaSO₄). Partial solvent evaporation and filtration through ashort silica gel path eluted with Et₂O and HPLC (silica gel;EtOAc:isooctane 4:96) gives 1.2a (3.27 g, 74%).

To a solution of 1.2a (1.3 g, 4.5 mmol) in MeOH (94 ml), is addeddropwise a 2M NaOH in MeOH (67 ml, 139.14 mmol). After 2 h solid CO₂ isadded and the solution is concentrated. The residue is poured into waterand extracted with Et₂O. The combined organic layers are washed withbrine, dried (NaSO₄) and after evaporation filtered through a short pathof silica gel, eluted with Et₂O. HPLC purification (silica gel;EtOAc:isooctane 4:96) gives 1.2b (1.25 g,94%).

Rf: 0.52 (isooctane:acetone 90:10).

IR (KBr): 1712, 1469, 1443, 1366, 1254 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 5.34 (1H, b s); 3.91 (1H, s); 2.83 (1H, td,J=5.81, 14.1); 2.58 (1H, dt, J=19.2, 26.45); 2.33 (1H, tm, J=15.07);2.22 (1H, ddd, J=13.9, 4.25, 2.4); 2.17 (1H, m); 2.09 (1H, dddd, J=13.9,5.9, 3.4, 2.5); 1.84 (1H, ddd, J=1.18, 4.28,14.1); 1.8 (1H, m); 1.74(1H, dd, J=16.6, 5.0); 0.94 (9H, s); 0.12 (6H, s); 0.65 (3H, s) ppm.

EXAMPLE 2

Synthesis of 1.3b

To a solution of 1.2b (1.1 g, 3.74 mmol) in MeOH (60 ml) solid NaBH₄(0.7 g, 18.67 mmol) is added in small portions at 0° C. The solution wasstirred overnight at r.t. The solution was concentrated and the residuewas dissolved in water. Extraction with CH₂Cl₂, washing of the combinedorganic layers with brine, drying (MgSO₄) solvent evaporation and HPLCpurification (silica gel; acetone:hexane 8:2) gives 1.3a (954 g, 86%with Rf 0.36; acetone:hexane 1:9).

To a cooled (0° C.) solution of 1.3a (800 mg, 2.7 mmol) and DIPEA (6 ml,65.61 mmol) in CH₂Cl₂ (30 ml), is added dropwise MEMCl (2.75 ml, 24.08mmol). The solution is stirred during 3 h at r.t. and is then dilutedwith Et₂O (100 ml). The mixture is washed with a 0.1 N HCl solution (30ml), saturated NaHCO₃ (30 ml) and brine. The organic layer is dried(MgSO₄) and concentrated. Column chromatography (silica gel;acetone:hexane 1:9), affords 1.3b (1.02 g, 98%).

Rf: 0.72 (acetone:hexane 1:9).

¹H NMR: (360 MHz, CDCl₃): δ: 5.3 (1H, m); 4.86 (1H, d, J=7.12); 4.69(1H, d, J=7.12); 3.67-3.79 (3H, m); 3.55 (2H,t,J=7.3); 3.38 (3H, s);3.22 (1H, dt, J=4.75, 10.1); 2.4-2.48 (1 H, m); 2.16 (1H,t, J=13.5);1.84-1.60 (5H, m); 1.63 (3H, s); 1.47-1.37 (3H, m); 0.9 (9H, s); 0.05(6H, s) ppm.

EXAMPLE 3

Synthesis of 1.4

To a solution of 1.3b (1 g, 2.6 mmol) in acetone:water 3:1 (20 ml), isadded NMMO (335 mg, 2.9 ml) and OsO₄ (100 mg, 0.39 mmol) and thesolution is stirred overnight at r.t. Solid Na₂S₂O₃ is added and themixture is extracted with CH₂Cl₂. The organic layers are dried (MgSO4),concentrated and filtered through a short path of silica gel (elutedwith Et₂O). The α-diol (86%) is dissolved in acetone:water 3:1 (24 ml)and the solution is cooled (0° C.). NalO₄ (1.132 g, 5.294 mmol) is addedin small portions to the solution. The mixture was stirred overnightunder Ar at r.t. The solution was then filtered, concentrated and theresidue taken up in water. The solution is extracted with CH₂Cl₂ (3×),the organic layers are washed with brine and dried (MgSO₄).

After evaporation of the solvent, a colourless oil is obtained (98%). It(650 mg, 1.56 mmol) is dissolved in a degassed solution of KOH (2 g) inwater (100 ml). The mixture is stirred overnight at 60° C.-80° C. underAr-flow. The solution was then extracted with CH₂Cl₂ and the organiclayers dried (Na₂SO₄). Filtration through a short path of silica gel(eluted with Et₂O) and HPLC purification (silica gel; acetone:hexane5:95), affords 1.4 (436 mg, 53%).

Rf: 0.81 (EtOAc).

IR (film): 1669; 1560; 1458; 1376; 1235; 1035 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.70 (1H, dd, J=5.25, 2.40); 4.83 (1H, d,J=7.07); 4.73 (1H, d, J=7.07); 4.72 (1H, m); 3.76-3.67 (2H, m); 3.64(1H, dt, J=10.36, 4.35); 3.55 (2H, dt, J=4.7); 3.38 (3H, s); 2.64 (1H,ddd, J=16.8, 6.6, 3.2); 2.45 (1H, ddd, J=22.4, 6.6); 2.35 (1H, dm,J=12); 2.24 (3H, s); 2.02 (1H, dddd, J=2.02, 3.98, 11.3, 16.8); 1.87(1H, m); 1.73 (1H, ddd, J=13.3, 6.03, 2.9); 1.60-1.43 (2H, m); 0.80 (9H,s); 0.01 (3H, s); −0.1 (3H, s) ppm.

EXAMPLE 4

Synthesis of 1.5a

A cooled (0° C.) solution of 1.4 (320 mg, 0.80 mmol) in hexane (3 ml),is stirred for 1.5 hour under 1 atm H₂ in the presence of 10% Pd/C (20mg) and is then filtered through a short path of silica gel (eluted withEt₂O). The solution is evaporated and the residue is dissolved in MeOH(10 ml). NaOMe (40 mg) is added at 0° C. and the solution is stirred for3 h while the temperature raised to r.t. The mixture is concentrated andthe residue is dissolved in saturated NH₄Cl solution. Extraction withEt₂O, drying (MgSO₄) and HPLC purification (silica gel; acetone:hexane1:9), affords 1.5a (311 mg, 97%).

Rf: 0.15 (acetone:petr.ether 1:9).

IR (film) 1709; 1471; 1357; 1253; 1201 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.77 (1H, d, J=7.08); 4.64 (1H, d, J=7.09);3.88 (1H, ppd, J=2.46); 3.68-3.59 (2H, m); 4.49 (2H, t, J=4.7); 3.32(3H, s); 3.31 (1H, dt, J=4.3, 10.5); 2.77 (1H, dt, J=10.6, 8.0); 2.06(3H, s); 1.99-1.89 (2H, m); 1.88-1.80 (1H, m); 1.76 (1H, ddd, J=11.4,3.7, 7.1); 1.69 (1H, ddd, J=13.7, 6.1, 3.0); 1.62-1.52 (2H, m);1.51-1.43 (1H, m); 1.38 (1H, ddt, J=13.7, 3.7, 2.3); 1.24-1.17 (1H, m);0.82 (9H, s); −0.05 (3H, s); −0.10 (3H, s) ppm.

EXAMPLE 5

Synthesis of 1.6a

To a cooled (0° C.) solution of Ph₃P+CH₃Br⁻ (112 mg, 0.31 mmol) in THF(1 ml) and HMPA (1 ml) under Ar, is added BuLi in THF (0.116 ml, 0.289mmol) followed after 1 h by 1.5a (48 mg, 0.12 mmol) in THF (1 ml). Thesolution is stirred for 3 h under Ar, while the temperature raised tor.t. Concentration and column chromatographic purification (silica gel;acetone:hexane 1:9), gives 1.5b (48 mg, 100% with Rf 0.47;acetone:hexane 1:9).

To a stirred solution of 1.5b (48 mg, 0.12 mmol) in THF (0.5 ml) underAr at r.t. is added a 0.5 M solution of 9-BBN in THF (1 ml, 0.5 mmol).After 4 h EtOH (0.1 ml), NaOH 6N (0.125 ml) and 30% H₂O₂ (0.25 ml) areadded, and the solution stirred under Ar for 1 h at 60° C. The mixtureis poured into brine and extracted with Et₂O (3×). Drying (MgSO₄),solvent evaporation and HPLC purification (silica gel; acetone:hexane15:85) gives 1.6a (37 mg, 74%).

Rf: 0.24 (acetone:hexane 2:8).

IR (film): 3445; 2930; 2878; 1469; 1364; 1252 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.82 (1H, d, J=7.1); 4.71 (1H, d, J=7.1);3.99 (1H, s); 3.75-3.63 (3H, m); 3.54 (2H, t, J=4.64); 3.4-3.32 (2H, m);3.38 (3H, s); 1.95-1.15 (12H, m); 1.10 (1H, t, J=9.81); 0.97 (3H, d,J=6.85); 0.88 (9H, s); 0.04 (3H, s); 0.02 (3H, s) ppm.

EXAMPLE 6

Synthesis of 1.8a

To a stirred solution at 0° C. of the alcohol 1.6a (35 mg, 0.083 mmol)in CH₂Cl₂ (2 ml) and Et₃N (0.5 ml), is added TsCl (32 mg, 0.168 mmol) at0° C. and the solution is stirred 12 h at r.t. The mixture is filteredthrough a short path of silica gel (eluted with acetone:hexane 15:85).HPLC purification (silica gel; acetone:hexane 15:85) gives 1.6b (43 mg,91% with Rf 0.26; acetone:hexane 15:85).

NaH (15 mg, 0.38 mmol) in DMSO (1.5 ml) is stirred for 2 h at 60° C.under Ar, the solution was then stirred at r.t.2-(1-ethoxy)-ethyloxy-2-methyl-3-butyne (547 mg, 3.5 mmol) is then addeddropwise at r.t. After 30 min 1.6b (200 mg, 0.35 mmol) in DMSO (1.3 ml)is added and the mixture stirred for 1.5 h at r.t. The mixture was thenpoured in saturated NaHCO₃ solution and extracted with Et₂O. Drying(MgSO₄), solvent evaporation and HPLC purification (silica gel;acetone:hexane 1:9) gives 1.7 (136 mg, 70% with Rf 0.38; acetone:hexane1:9).

1.7 (40 mg, 0.0721 mmol) is dissolved in EtOAc (3 ml) and 10% Pd/C (3mg) is added. The suspension was shacked under 4 bar H2 for 1 h at r.t.The mixture is then filtered through a short path of silica gel (elutedwith EtOAc) giving 1.8a (12 mg, 34%).

Rf: 0.20 (acetone:hexane 15:85).

IR (film): 3456; 2932; 2861; 1368; 1251 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.83 (1H, d, J=7.1); 4.71 (1H, d, J=7.1);3.92 (1H, s); 3.76-3.68 (2H, m); 3.56 (2H, t, J=4.7); 3.39 (3H, s); 3.35(1H, dt, J=10.3, 4.0); 1.96-1.73 (5H, m); 1.61-1.22 (10H, m); 1.21 (6H,s); 1.14-0.98 (4H, m); 0.88 (12H, m); 0.03 (3H, s); 0.01 (3H, s) ppm.

EXAMPLE 7

Synthesis of 1.8d

To a cooled (−78° C.) and stirred solution of 1.8a (10 mg, 0.02054 mmol)in CH₂Cl₂ (1 ml), is added 1.5 M Me₂BBr in CH₂Cl₂ (0.034 ml, 0.05 mmol).The mixture is stirred for 3 h at −78° C. and is then added dropwise toa vigorously stirred solution of saturated NaHCO₃ and THF. The mixturewas extracted with Et₂O. Drying (MgSO₄), solvent evaporation and HPLCpurification (silica gel; acetone:hexane 2:8), gives 1.8b (6 mg, 73%).

To a solution of diol 1.8b (5 mg, 0.0125 mmol) in CH₂Cl₂ (2 ml) at r.t.is added PDC (14 mg, 0.0376 mmol). The solution is stirred for 4 h andis then filtered through a short path of silica gel (acetone:hexane 3:7)giving 1.8c (5 mg, 99%).

Rf: 0.21 (acetone:hexane 2:8).

¹H NMR: (500 MHz, CDCl₃): δ: 3.93 (1H, br s); 3.35 (1H, dt, J=4.0,10.1); 1.96-1.15 (17H, m); 1.21 (6H, s); 1.12-0.99 (3H, m); 0.9-0.88(12H, m); 0.05 (3H, s); 0.02 (3H, s) ppm.

The ketone 1.8c (5 mg, 0.0126 mmol) was dissolved in THF (1 ml) and TSIM(0.5 ml, 3.41 mmol) is added. The mixture is stirred for 1 h at r.t. andis then purified by chromatography (silica gel; acetone:hexane 1:9)giving 1.8d (5.8 mg, 98% with Rf 0.55; acetone:hexane 1:9).

EXAMPLE 8

Synthesis of 1.9c

Alkene 1.5b (105 mg, 0.264 mmol) is stirred with 1M TBAF (1 ml, 1 mmol)in THF (1.5 ml) for 10 d at 30° C. The mixture is concentrated andpurified by chromatography (silica gel; acetone:hexane 3:7) giving 1.9a(5.8 mg, 99%).

To a suspension of NaH (70 mg, 2.92 mmol) in THF (5 ml) is addeddropwise 1.9a (32 mg, 0.113 mmol) in THF (2 ml). After stirring for 0.5h and cooling (0° C.) CS₂ (0.349 ml, 5.8 mmol) is added dropwise and thestirring is continued for 24 h while the temperature raised to r.t. MeI(0.375 ml, 6 mmol) is added dropwise and the solution is stirred for 2h. The mixture is poured in a 0.1 N HCl solution. Extraction with Et₂O,drying (MgSO₄), evaporation and HPLC purification (silica gel;acetone:hexane 2:8), gives 1.9b (41 mg, 97%).

To a solution of Bu₃SnH (1 ml, 1.08 mmol) and AIBN (2 mg) in toluene (5ml) at 110° C., is added dropwise (0.5 h) 1.9b (41 mg, 0.11 mmol) intoluene (2 ml). The mixture is stirred for 8 h at 110° C. Columnchromatography (silica gel; acetone:hexane 1:9), followed by HPLC(silica gel; acetone:hexane 3:7), gives 1.9c (27 mg, 92%).

Rf: 0.3 (acetone:hexane 5:95).

IR (film): 2927; 2872; 1447; 1113; 1043 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.83 (1H, d, J=7.1); 4.71 (1H, d, J=7.1);4.7-4.67 (2H, m); 3.77-3.66 (2H, m); 3.57 (2H, t, J=4.8); 3.4 (3H, s);3.38 (1H, dt, J=4.2, 9.9); 2.16 (1H, m); 2.08 (1H, m); 1.99 (1H, m);1.88-1.76 (2H, m); 1.67 (3H, s); 1.57-1.46 (2H, m); 1.36-1.1 (6H, m);0.90-0.80 (2H, m) ppm.

EXAMPLE 9

Synthesis of 1.10b

From 1.9c as described for 1.6b from 1.5b. Overall yield of the epimericmixture 20-S, 20-R (ratio 8:2) is 82%.

Rf: 0.35 (acetone:hexane 15:85).

IR (film): 2931; 2876; 1458; 1362; 1189 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 7.78 (2H, d, J=8.3); 7.35 (2H, d, J=8.3);4.80 (1H, d, J=7.1); 4.68 (1H, d, J=7.1); 3.97 (1H, dd, J=9.5, 5.0);3.83 (1H, m); 3.73-3.64 (2H, m); 3.54 (3H, t, J=4.7); 3.39 (3H, s); 3.38(3H, s); 3.28 (1H, dt, J=10.1, 4.3); 2.54 (3H, s); 2.03 (1H, m);1.91-1.81 (2H, m); 1.78-1.47 (4H, m); 1.3-1.03 (5H, m); 0.95-0.79 (2H,m); 0.91 (3H, d, J=6.9); 0.78 (3H, d, J=6.9) ppm.

EXAMPLE 10

Synthesis of 1.10d

To a solution of 1.10b (40 mg, 0.091 mmol) in DMSO (1.5 ml), is added Kl(150 mg, 0.91 mmol). The mixture is stirred for 4 h at 60° C. and isthen poured out in brine. The solution was extracted. Extraction withEt₂O, drying (MgSO₄), solvent evaporation and flash chromatography gives1.10c (34.3 mg, 95%). 1.10c (5 mg, 0.0126 mmol) is dissolved inEtOH:water 7:3 (0.5 ml). Cul (20 mg, 0.105 mmol), Zn powder (30 mg,0.458 mmol) and methylvinylketone (0.150 ml, 1.81 mmol) are added andthe solution is stirred for 35 minutes at 15° C. in a sonoficator(Banson 220). The sonofication process is stopped in order to cool downthe liquid in the sonoficator and the process is repeated for 40minutes. Extra Cul (10 mg, 0.105 mmol), Zn (14 mg, 0.23 mmol) andmethylvinylketone (0.075 ml, 0.95 mmol) are added and the sonoficationprocess is continued for 2 h. The mixture is then filtered through ashort path of silica gel (eluted with Et₂O) and dried (MgSO₄). HPLCpurification (acetone:hexane 15:85), gives 1.10d (3.6 mg, 83%).

Rf: 0.18 (acetone:hexane 5:95).

IR (film): 2928; 2871; 1716; 1459 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.82 (1H, d, J=7.1); 4.7 (1H, d, J=7.1);3.96-3.66 (2H, m); 3.56 (2H, t, J=4.7); 3.39 (3H, s); 3.33 (1H, dt,J=10.4, 4.3); 2.44-2.34 (2H, m); 2.12 (3H, s); 2.05 (1H, dm, J=12.4);1.91 (1H, m); 1.79 (1H, dm, J=11.0); 1.7-1.4 (5H, m); 1.36-1.2 (4H, m);1.19-0.94 (4H, m); 0.88 (3H, d, J=8.5); 0.87 (1H, m); 0.77 (3H, d,J=6.6) ppm.

EXAMPLE 11

Synthesis of 1.11d

Ketone 1.10d (10 mg, 0.0294 mmol) is dissolved in THF (1.5 ml) and 2MMeMgCl (0.4 ml, 1.2 mmol) is added. The mixture is stirred for 1 h atr.t., 0.1 N HCl is then added until the formation of gas stopped. Thesolution is filtered through a short path of silica gel and anhydrousMgSO₄ giving 1.11a (10.3 mg, 98%).

A solution of 1.11a (10 mg, 0.028 mmol) in MeOH (2 ml) is stirredtogsilica gel with Amberlyst 15 (200 mg) for 1 week at 30° C. Themixture is then filtered through a short path of silica gel (eluted withEt₂O). HPLC purification (silica gel; acetone:hexane 3:7) gives 1.11b(7.2 mg, 96%).

Alcohol 1.11b is then transformed into 1.11d (Rf 0.58; acetone:hexane15:85) as described for 1.8d from 1.8b (80% yield).

Rf: 0.2 (acetone:hexane 15:85).

IR (film): 3422; 2958; 2872; 1713; 1464; 1377 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 2.37-2.22 (3H, m); 2.13 (1H, m); 2.07 (1H,dm, J=12.4); 1.74-1.20 (15H, m); 1.21 (6H, s); 1.09 (1H, m); 0.90 (3H,d,J=6.83); 0.78 (3H, d, J=6.78) ppm.

EXAMPLE 12

Synthesis of Acid 2.1

A suspension of benzeneselenic acid (9.6 g, 0.05 mol) in a mixture ofTHF (50 ml) and phosphate buffer (0.1M, pH=7, 25 ml) was treated withca. 30% hydrogen peroxide (88 g, 0.4 mol) at room temperature. Asolution of menthone (6.16 g, 0.04 mol) in THF (25 ml) was added and thereaction mixture was stirred at room temperature for 17 h. SaturatedNaHCO₃ aq. solution was added until the pH of the reaction mixturereached 9. After removal of H₂O₂ and THF under reduced pressure, thereaction mixture was acidified to pH 5. After saturation with salt, thereaction mixture was extracted with ether (250 ml, 3 times) and thecombined ether phases were dried over anhydrous MgSO₄. After filtration,the filtrate was concentrated in vacuo. The remaining colourless liquid(14 g) was dissolved in 150 ml of methanol and 37% HCl (3.75 ml) wasadded. This mixture was refluxed for 3 hours. After cooling, thereaction mixture was treated with saturated NaHCO₃ aq. solution to pH 8.The organic solvent was removed by evaporation under reduced pressure.The remaining residue was extracted with ether (3 times). The combinedether solution was dried over anhydrous MgSO₄. After filtration andconcentration the crude material was purified by column chromatography(EtOAc/hexane 1:4) providing pure methyl ester (7.32 g, 91%).

Rf: 0.45 (EtOAc:hexane 1:2).

IR (film): 3434 (m); 2957, 2873 (s); 1736 (s); 1461, 1437 (m); 1287,1261, 1205, 1164 (s); 734 (s) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 3.68 (3H, s); 2.34 (1H, dd, J=6.1, 14.8);2.14 (1H, dd, J=8.0, 14.8); 1.95 (1H, m); 1.65 (1H, m); 1.52 (2H, m);1.35 (2H, m); 1.22 (1H, m); 0.96 (3H, d, J=6.8); 0.90 (6H, dd, J=7.1,7.5) ppm.

MS (m/z): 202 (2%); 187 (1%); 184 (2%); 159 (2%); 43 (100%).

To a solution of the previous ester (5.17 g, 25.5 mmol) in DMF,tert-butyldimethylsilyl chloride (RBDMS-CI, 5.79 g, 38.4 mmol), DMAP (50mg) and imidazole (3.92 g, 57.6 mmol) were added. The solution wasstirred overnight at room temperature under nitrogen atmosphere. Dilutedwith ether, the reaction mixture was washed with water. The organicphase was dried over anhydrous MgSO₄. After filtration andconcentration, the remaining crude material was purified by columnchromatography (silica gel, EtOAc:hexane 1:50), yielding 7.85 g ofproduct (98% yield).

Rf: 0.58 (EtOAc:hexane 1:2).

IR (film): 2896 (s); 2857 (s); 1743 (s); 1471; 1462, 1436, 1385 (m);1253 (s); 1210, 1165, 1101 (m); 1057, 837, 773 cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 3.67 (3H, s); 3.40 (1H, m); 2.30 (1H, dd,J=6.4, 14.8); 2.12 (1H, dd, J=8.0, 14.8); 1.90 (1H, m); 1.68 (1H, m);1.40 (3H, m); 1.15 (1H, m); 0.95 (3H, d, J=6.8); 0.88 (9H, s); 0.84 (6H,dd, J=6.8, 10.5); 0.02 (6H, s) ppm.

MS (m/z): 316 (1%); 301 (2%); 249 (3%); 191 (5%); 115 (80%).

To a stirred suspension of potassium tert-butoxide (16.85 g, 165 mmol)in dry diethylether (150 ml) was added 0.752 ml of water via syringe at0° C. The resulting slurry was stirred for 10 minutes at the sametemperature and was then treated with the previous product (6 g, 19mmol). The ice bath was removed and the reaction mixture was stirred atroom temperature for 50 hours. To the reaction mixture ice was addeduntil two clear layers are formed. This mixture was acidified with 10%HCl aq. solution till pH=1. After extraction with ether the combinedether phases were dried over MgSO₄. Following filtration andconcentration, the crude product was purified by silica gel columnchromatography 1:20 and 1:4 EtOAc:hexane) affording 5.34 g of acid 2.1(yield: 93%).

Rf: 0.54 (EtOAc:hexane 1:″).

IR (film): 3500 (m); 2958, 2857 (s); 1708 (s); 1471, 1462, 1410 (s);1294, 1252,1226 (s) 836, 773 (s) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 3.40 (1H, m); 2.35 (1H, dd, J=6.0, 15.0Hz); 2.15 (1H, dd, J=8.0, 15.0 Hz); 1.92 (1H, m); 1.70 (1H, m); 1.42(4H, m); 1.00 (3H, d, J=6.8); 0.88 (9H, s); 0.87 (6H, dd, J=6.6, 10.5);0.03 (6H, s) ppm.

MS (m/z) 302 (1%); 287 (1%); 258 (10%); 245 (5%); 187 (50%); 115 (80%).

EXAMPLE 13

Synthesis of Ester 2.2

To a stirred solution of (R)-3-methyl-2-cyclohexen-1-ol (0.70 g, 6.25mmol) in methylene chloride (50 ml) was added acid 2.1 (1.51 g, 5 mmol)at 0° C. After addition of DCC (3.25 g, 15.8 mmol) and DMAP (0.732 g, 6mmol) at the same temperature, the mixture was kept for 5 minutes at 0°C. and was then warmed till room temperature and allowed to be stirredat r.t. overnight. 2 ml of ethanol and acetic acid were addedrespectively and the mixture was further stirred at r.t. for 2 h. Afterfiltration, the reaction mixture was concentrated till 20 ml. Afterdilution with diethyl ether (200 ml), the reaction mixture was washedwith water. The ether solution was dried over anhydrous MgSO₄ anddoncentrated in vacuo. The residual liquid was separated by silica gelcolumn chromatography affording 1.8 g of ester 2.2 (yield: 91%).

Rf: 0.6 (EtOAc:hexane 1:20).

IR (film): 2950 (s); 2857 (s); 1730 (s); 1462, 1380 (m); 1251 (s): 1162,1055 (m) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 5.45 (1H, m); 5.25 (1H, m); 3.40 (1H, m);2.30 (1H, dd, J=8,14); 2.10 (1H, dd, J=8, 15); 1.95 (2H, m); 1.75 (3H,m); 1.70 (3H, s); 1.65 (2H, m); 1.62 (2H, m); 1.40 (4H, m); 1.10 (1H,m); 0.95 (3H, d, J=6.4); 0.88 (9H, s); 0.85 (6H, q, J=7, 14); 0.02 (6H,s) ppm.

MS (m/z): 396 (1%); 339 (1%); 267 (1%); 167 (30%); 109 (50%); 95 (80%);75 (100%).

EXAMPLE 14

Synthesis of Acid 2.3

To a stirred solution of N-isopropyl-N-cyclohexylamine (158 mg, 1.12mmol) in 1 ml of dry hexane, n-butyllithium (2.40M solution in hexane,0.467 ml, 1.12 mmol) was added dropwise at −5° C. over several minutes.Following the addition, the colourless solution was stirred at −5° C.for 20 minutes. After which the hexane and excess amine was removedunder vacuo at 0° C. Under argon the residual white solid was dissolvedin THF (2 ml) and HMPA (0.7 ml). The mixture was cooled to −78° C. andacid 2.2 was added dropwise over 2 minutes. After 10 min following theaddition, the reaction mixture was allowed to warm until −30° C. and waskept at this temperature for 1 h. The reaction mixture was cooled to−78° C. and TBDMS-CI (168 mg, 1.12 mmol) was added. The reaction mixturewas stirred at −78° C. for 10 min then was warmed to room temperaturevery slowly within 1 hour. Finally the reaction mixture was refluxedunder argon for 17 hours and was then cooled down to room temperature.After dilution with ether, the reaction mixture was washed with 2.5% HClaq. solution and water. The organic phase was dried over anhydrous MgSO₄and concentrated in vacuo. The residue was separated by silica gelcolumn chromatography (EtOAc:hexane) affording 117 mg of acid 2.3 and220 mg of starting material 2.2 (yield: 67% based on consumed startingmaterial).

Rf: 0.56 (EtOAc:hexane 1:5).

IR (film): 3400 (m); 2980 (s); 1704 (s); 1462, 1381 (m); 1253, 1202 (m)cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 5.26 (1H, m); 5.40 (1H, d, J=10.3); 3.36(1H, m); 2.21 (1H, d, J=5.5); 1.95 (2H, m); 1.60-1.80 (6H, m); 1.42 (1H,s); 1.25 (3H, m); 1.11 (3H, s); 1.02 (3H, d, J=6.8 Hz); 0.88 (9H, s);0.83 (3H, d, J=6.8); 0.82 (3H, d, J=6.8); 0.03 (6H, s) ppm.

MS (m/z): 396 (1%); 352 (1%); 381 (1%); 339 (1%); 281 (2%); 237 (5%);115 (30%); 95 (80%).

EXAMPLE 15

Synthesis of Alkene 2.4

To a solution of acid 2.3 (474 mg, 1.2 mmol) in dry ether (10 ml) wasadded 30 ml of diazomethane (0.5M solution in ether) at 0° C. Thereaction mixture was stirred at 0° C. for 1 hour. The ether and excessdiazomethane were removed by evaporation under reduced pressure. Theresidue (425 mg, 86% yield) was dissolved in THF and added to asuspension of lithium aluminum hydride (114 mg, 3 mmol) in THF (20 ml)by syring. The reaction mixture was stirred at room temperature for 3hours and was then refluxed for 1 hour. Excess lithium aluminum hydridewas destroyed by careful addition of ethanol and was then treated withdiluted HCl aq. solution. The alcohol was extracted with ether and thecombined ether phases were dried over anhydrous MgSO₄. Crude product,after filtration and concentration, was isolated by silica gel columnchromatography and purified by HPLC yielding 382 mg of alcohol (89%yield).

Rf: 0.4 (EtOAc:hexane 1:10).

IR (film): 3355 (m); 2956 (s); 1462, 1385, 1251, 1048 (s); 836, 773 (s);941, 732, 664 (m) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 5.67 (1H, m); 5.53 (1H, d, br, J=10); 3.81(1H, q, J=6.0, 11.4); 3.72 (1H, dd, J=5.5, 11.3); 3.38 (1H, m); 1.95(2H, m); 1.70 (2H, m); 1.60 (4H, m); 1.40 (1H, m); 1.35-1.15 (4H, m);1.03 (3H, s); 1.00 (3H, s); 0.90 (9H, s); 0.85 (6H, dd, J=6.9, 9.9);0.05 (6H, s) ppm.

MS (m/z): 339 (M.+ -iPr, 1%); 341 (1%); 325 (1%); 251 (1%).

A solution of the alcohol (250 mg, 0.65 mmol) in pyridine (10 ml) wasadded p-toluenesulphonylchloride (420 mg, 2.2 mmol) at room temperature.The light yellow solution was stirred at room temperature for 18 hours,then was poured onto ice. This mixture was extracted with ether and thecombined ether phases were washed with 5% HCl aq. solution until pH=3.After drying over anhydrous MgSO₄, the organic phase was concentrated invacuo. The residue was filtered through a short silica gel column andpurified by HPLC giving 336 mg of tosylate (yield: 96%).

Rf: 0.37 (EtOAc:hexane 1:20).

IR (film): 2958, 2857 (s); 1741, 1599 (m); 1462, 1367 (s); 1250, 1178(s); 1047, 953 (s); 837, 773 (s) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 7.80 (2H, d, J=8.3); 7.33 (2H, d, J=8.4Hz); 5.59 (1H, m); 5.32 (1H, d, br, J=10.2); 4.20 (1H, dd, J=4.7, 10.0);4.10 (1H, dd, J=7.4, 10.0); 3.30 (1H, m); 2.46 (3H, s); 1.90 (2H, m);1.62 (2H, m); 1.53 (3H, m); 1.45 (3H, m); 1.26 (2H, m); 1.15 (1H, m);0.95 (3H, s); 0.90 (3H, d, J=7.1); 0.88 (9H, s); 0.82 (6H, dd, J=6.9,8.8); 0.01 (3H, s); −0.01 (3H, s) ppm.

MS (m/z): 512 (1%); 486 (1%); 455 (1%); 426 (1%); 364 (2%); 321 (2%);307 (5%); 229 (20%); 9.5 5 (100%).

To a suspension of lithium aluminum hydride (71 mg, 1.88 mmol) in THF(12 ml) was added the tosylate (336 mg, 0.627 mmol) as a solution in THFat r.t.

The reaction mixture was refluxed for 2 hours. Excess LiAlH₄ wasdestroyed by adding ethanol. The mixture was then treated with 5% HClaq. solution. This mixture was extracted with diethyl ether and thecombined ether phases were dried over anhydrous MgSO₄. Pure alkene 2.4(234 mg) was isolated by column chromatography (fine silica gle) in 91%yield.

Rf: 0.54 (pure hexane).

IR (film): 3011 (w); 2957, 2858 s); 1462, 1383, 1386 (s); 1253, 1082,1054 (s); 836, 772 (s); 941, 731 (w) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 5.58 (1H, m); 5.45 (1H, m); 3.37 (1H, m);1.92 (2H, m); 1.70 (1H, m); 1.60 (4H, m); 1.52 (2H, m); 1.36 (1H, m);1.28 (1H, m); 1.20 (1H, m); 0.93 (3H, s); 0.89 (9H, s); 0.87 (3H, d,J=6.8); 0.85 (3H, d, J=7;3); 0.82 (6H, q, J=6.9); 0.02 (3H, s); 0.01(3H, s) ppm.

MS (m/z): 366 (1%); 364 (1%); 309 (10%); 287 (1%); 233 (5%); 75 (100%).

EXAMPLE 16

Synthesis of Cyclohexanone 2.5

To a solution of alkene 2.4 (60 mg, 0.164 mg) in THF (6 ml) was added9-BBN (0.5M solution in THF, 3.3 ml, 1.04 mmol) at room temperatureunder nitrogen atmosphere. The solution was stirred at room temperaturefor 1 hour and was then refluxed for 20 hours. The organoborane wasoxidized by adding, successively ethanol (0.5 ml), 6N NaOH (0.4 ml) and30% hydrogen peroxide (0.8 ml). This mixture was heated at 50° C. for 1hour. The reaction mixture was extracted with ether and the combinedether phases were washed with 5% HCl aq. solution. The organic phase wasdried over anhydrous MgSO₄, filtered and the filtrate was concentratedin vacuo. The residue was purified by column chromatography (fine silicagel) affording the corresponding alcohol (51 mg) in 80% yield as amixture of diastereomers.

A mixture of this alcohol (51 mg, 0.133 mmol) and PDC (175 mg, 0.442mmol) in methylene chloride (4 ml) was stirred at room temperature for15 hours and was directly purified on silica gel. Final purification byHPLC led to ketone 2.5 (46 mg) in 90% yield.

Rf: 0.4 (EtOAc:hexane 1:10).

IR (film): 2931, 2857 (s); 1715 (s); 1472, 1385 (s); 1250, 1081, 1058(s); 941, 667 (m); 837, 773 (s) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 3.36 (1H, m); 2.26 (3H, m); 2.05 (1H, d,J=13.3); 1.90 (1H, m); 1.82 (1H, m); 1.65 (3H, m); 1.58 (1H, m); 1.50(3H, m); 1.25 (1H, m); 0.92 (3H, d, J=6.9); 0.88 (9H, s); 0.85 (3H, s);0.84 (6H, q); 0.79 (3H, d, J=7;3); 0.02 (6H, s) ppm.

MS (m/z): 382 (2%); 368 (10%); 340 (1%5); 326 (60%); 185 (60%); 95(70%); 75 (100%).

EXAMPLE 17

Synthesis of Alkene 2.6

A solution of 2.4 (160 mg, 0.437 mmol) and TBAF (1M solution in THF,2.18 ml, 2.18 mmol) in THF (10 ml) was heated at 30° C. with stirringfor 3 days. The reaction mixture was diluted with hexane and wasimmediately chromatographed. The reaction mixture was diluted withhexane and was immediately chromatographed. The crude product wasfurther purified by HPLC (1:12 EtOAc/hexane) to afford the unprotected24S-alcohol (106 mg, 88%).

IR (film): 3378 (m); 2959, 2870 (s); 1646 (w); 1462, 1380 (s); 1060, 989(m); 732 (m) cm⁻¹.

¹H NMR (500 MHz, CDCl₃): δ: 5.59 (1H, dt, J=10.2, 3.5); 5.46 (1H, dq,J=10.2, 2.0); 3.31 (1H, m); 1.92 (2H, m); 1.65 (1H, m); 1.58 (4H, m);1.37 (2H, m); 1.30-1.20 (4H, m); 0.94 (3H, s); 0.92 (3H, d, J=6.8); 0.90(3H, d, J=6.8); 0.88 (3H, d, J=6.8); 0.82 (3H, d, J=7.3).

MS (m/z): 252 (M.⁺, 3); 234 (5); 149 (20); 122 (20); 95 (100) ppm.

A solution of the above alcohol (100 mg, 0.397 mmol) in THF (4 ml) wastreated with triphenylphosphine (260 mg, 0.99 mmol) and 4-nitrobenzoicacid (166 mg, 0.99 mmol) under nitrogen atmosphere at r.t. Diethylazodicarboxylate was subsequently slowly added. The reaction mixture wasstirred at r.t. for 15 hours. After dilution with hexane the mixture wasthen filtered through a silica gel column. The further purification withHPLC afforded the corresponding inverted p-nitrobenzoate ester (100 mg,63%).

A mixture of the latter (100 mg, 0.25 mmol) and K₂CO₃ (173 mg, 1.25mmol) in methanol was stirred at room temperature for 0.5 h. No reactionwas detected. To the reaction mixture was then added KOH (745 mg) andthe mixture was stirred at room temperature for 1.5 h. Water was addedand the mixture was extracted with ether. The combined ether solutionwas washed with water, dried over anhydrous MgSO₄, and concentrated invacuo. The crude material was purified by HPLC (1:11 EtOAc/hexane) togive the 24R-alcohol (59 mg, 94%).

IR (film): 3379 (m); 2959, 2870 (s); 1644 (w); 1462, 1380 (s); 1060, 989(m); 732 (m) cm⁻¹.

¹H NMR (500 MHz, CDCl₃) : δ5.59 (1H, ddd, J=3.4, 4.2, 10.2 Hz); 5.46(1H, dq, J=10.1, 2.8); 3.3 (1H, m); 1.92 (2H, ); 1.70 (1H, m); 1.63 (1H,m); 1.58 (3H, m); 1.46 (2H, m); 1.35 (1H, m); 1.28 (2H, m); 1.10 (1H,m); 0.94 (3H, s); 0.92 (3H, d, J=6.7 Hz); 0.90 (3H, d, J=6.7); 0.87 (3H,d, J=0.87); 0.82 (3H, d, J=7.3). ppm

MS (m/z): 252 (M.+, 3); 234 (5); 149 (20); 122 (20); 95 (100).

A solution of this alcohol (59 mg, 0.234 mmol), imidazole (32 mg, 0.468mmol), TBDMS-CI (71 mg, 0.468 mmol) and DMAP (10 mg) in DMF (3 ml) wasstirred at room temperature for 16 hours, and was then treated withTBDMS-CI (71 mg, 0.468 mmol), imidazole (32 mg), and DMAP (10 mg). After5 hours stirring at room temperature the addition was repeated oncemore. The reaction mixture was stirred 10 hours and was then treatedwith 10% HCl (1 ml). After 10 min stirring the reaction mixture wasextracted with ether. The combined ether solution was washed with 5% HCland water, dried over MgSO₄ and concentrated in vacuo. The residualmaterial was separated by column chromatography and purified by HPLC(pure hexane) to give 2.6 (83 mg, 97%).

IR (film): 2857 (s); 1645 (w); 1408; 1375 (m); 1289, 1156 (m); 945 cm⁻¹.

¹H NMR (500 MHz, CDCl₃): δ5.58 (1H, dt, J=10.2, 3.4); 5.45 (1H, dq,J=10.1, 1.8); 3.36 (1H, q, J=5.0); 1.91 (2H, m); 1.68 (1H, m); 1.58 (4H,m); 1.48 (1H, m); 1.36 (2H, m); 1.27 (2H, m); 1.22 (1H, m); 0.93 (3H,s); 0.89 (9H, s); 0.87 (3H, d, J=6.8); 0.84 (3H, d, J=6.8); 0.83 (3H, d,J=6.8); 0.80 (3H, d, J=7.3); 0.03 (3H, s); 0.02 (3H, s) ppm

MS (m/z): 366 (M.⁺, 1%).

EXAMPLE 18

Synthesis of Cyclohexanone 2.7

To a solution of 2.6 (80 mg, 0.219 mmol) in THF (8 ml) was added 9-BBN(0.5 M solution in THF, 4.37 ml, 2.19 mmol) at r.t. under nitrogenatmosphere. The reaction mixture was refluxed for 20 hours. Theorganoboranes were oxidized by adding, successively EtOH (0.66 ml), 6NNaOH (0.53 ml) and 30% H₂O₂ (1.06 ml). This mixture was heated at 50° C.for 1 hour. After dilution with ether the reaction mixture was washedwith 5% HCl aq. solution, water and dried over MgSO₄. Afterconcentration the residual oil was chromatographed and was furtherpurified by HPLC to give the alcohol (77.3 mg, 92%).

A solution of the latter (77.3 mg, 0.2 mmol) and PDC (396 mg, 1 mmol) inCH₂Cl₂ (10 ml) was stirred at room temperature for 24 hours. Directcolumn chromatography of the reaction mixture followed by HPLCpurification afforded the desired ketone 2.7 (71 mg, 92%).

¹H NMR (500 MHz, CDCl₃): δ3.36 (1H, dt, J=9.7, 4.8); 2.28 (1H, d,J=13.7); 2.26 (2H, m); 2.05 (1H, dt, J=13.4, 1.4); 1.90 (1H, m); 1.82(1H, m); 1.70-1.60 (4H, m); 1.50 (1H, m); 1.34-1.20 (3H, m); 0.97 (1H,m); 0.89 (3H, d, J=6.8); 0.88 (9H, s); 0.86 (3H, s); 0.84 (3H, d,J=6.8); 0.83 (3H, d, J=6.8); 0.79 (3H, d, J=7.2); 0.02 (3H, s); 0.01(3H, s) ppm.

EXAMPLE 19

Synthesis of Ester 2.9

To a stirred solution of (R)-(+)-citronellic acid (2.8; 0.98 g, 5.76mmol) in methylene chloride (50 ml) was added(R)-3-methylcyclohexen-1-ol (84% e.e., 0.64 g, 5.76 mmol) at 0° C. undernitrogen atmosphere. The reaction was initiated by the addition of DCC(2.96 g, 14.4 mmol) and DMAP (0.732 g, 6 mmol). After 5 min at 0° C. thereaction mixture was warmed to room temperature and allowed to stir atr.t. overnight (18 h). Ethanol (4 ml) and acetic acid (4 ml) were addedto the reaction mixture at 0° C. The mixture was stirred at 0° C. for 20min and at room temperature for 1 hour. Ether was added and the formedwhite solid was removed by filtration. The filtrate was evaporated underreduced pressure and the residual liquid was dissolved in ether. Theether solution was washed with water, dried over anhydrous MgSO₄. Columnchromatography of the crude material afforded ester 2.9 (1.521 g) in 96%yield.

Rf: 0.52 (EtOAc:hexane 1:20).

IR (film): 2931 (s); 2360 (w); 1732 (s); 1456, 1378 (m); 1150, 1071 (s);921 (m) cm⁻¹.

¹H NMR (360 MHz, CDCl₃): δ5.45 (1H, m); 5.25 (1H, m); 5.07 (1H, t, J=7.1Hz); 2.28 (1H, dd, J=6.0, 14.4 Hz); 2.10 (1H, dd, J=8.2, 14.4); 1.95(3H, m); 1.71 (3H, s); 1.68 (3H, s); 1.58 (3H, s); 1.70 (4H, m); 1.20(4H, m); 0.92 (3H, d, J=6.6) ppm.

MS (m/z): 264 (M.⁺, 1%); 249 (1%); 227 (1%); 191 (1); 169 (30); 109(30); 95 (100).

EXAMPLE 20

Synthesis of Acid 2.10

To a stirred solution of diisopropylamine (456 μl, 3.27 mmol) in THF (10ml) was added n-butyllithium (2.45M solution in hexane, 1.33 ml, 3.27mmol) at −15° C. under nitrogen atmosphere. The reaction mixture wasstirred at the same temperature for 20 min and then HMPA (3 ml) wasadded. The reaction mixture was cooled to −78° C. A solution of 2.9(0.77 g, 2.92 mmol) in THF (2 ml) was added 2.9 to the reaction mixturevery slowly at −78° C. After 10 min following the addition, the formedenolate is allowed to warm to −50° C. for 20 min. TBDMS-CI (491 mg, 3.27mmol) as solid was added at −50° C. and the reaction mixture was stirredat the same temperature for 20 min and was then warmed to roomtemperature. The reaction mixture was stirred at room temperature for 3hours and was then refluxed for 16 hours. 5% HCl aq. solution (15 ml)was added and the mixture was stirred at room temperature for 60 min.The mixture was extracted with ether. The combined ether solution waswashed with water, dried over anhydrous MgSO₄ and concentrated in vacuo.The residual oil was separated by column chromatography to afford 2.10(448 mg, 58%).

Rf: 0.35 (EtOAc/hexane 1:5).

IR (film): 2930 (s); 1704 (s); 1462, 1381 (m); 1285, 1253, 1202 (m); 836(m) cm⁻¹.

¹H NMR (360 MHz, CDCl₃): δ5.62 (1H, m); 5.41 (1H, d, J=11.0); 5.09 (1H,t, j=6.9 Hz); 2.22 (1H, d, J=5.9); 2.08 (1H, m); 1.90 (3H, m); 1.68 (3H,s); 1.58 (3H, s); 1.62 (7H, m); 1.10 (3H, s); 1.02 (3H, d, J=6.8) ppm.

MS (m/z): 264 (M.⁺, 5%); 249 (1%); 221 (1); 208 (5); 154 (15); 109 (15);96 (100).

EXAMPLE 21

Synthesis of Alkene 2.11

To a suspension of LiAlH₄ (302 mg, 7.95 mmol) in THF (10 ml) was added asolution of 2.10 (420 mg, 1.59 mmol) in THF (5 ml). The reaction mixturewas refluxed for 48 hours. The excess LiAlH₄ was destroyed by additionof 5% HCl aq. solution. The mixture was extracted with ether. Thecombined ether phases were washed with water, dried over MgSO₄ andconcentrated in vacuo. The residual material was chromatographed to givethe primary alcohol (344 mg, 88%). A diastereoisomer could be removed byfurther HPLC purification.(EtOAclhexane 1:6).

Rf: 0.35 (EtOAc/hexane 1:5).

IR (film): 3339 (m); 2928, 2871 (s); 1454, 1376 (m); 1028 (m); 732 (m)cm⁻¹.

¹H NMR (500 MHz, CDCl₃): δ5.67 (1H, dt, J=10.1, 3.5 Hz); 5.53 (1H, dt,J=10.2, 1.8 Hz); 5.12 (1H, tt, J=6.8, 1.6); 3.78 (1H, m); 3.72 (1H, m);2.06 (1H, m); 1.93 (2H, m); 1.88 (1H, tt, J=7.8. 7.8); 1.70 (2H, m);1.68 (3H, s); 1.61 (2H, m); 1.59 (3H, s); 1.50 (1H, m); 1.40 (1H, ddd,J=1.2, 4.5, 10.5); 1.33 (1H, m); 1.28 (2H, m); 1.13 (1H, m); 1.13 (3H,d, J=7.0); 1.01 (3H, s) ppm.

MS (m/z): 250 (M.⁺, 5); 232 3); 219 (10); 137 (40); 95 (100).

To a solution of the alcohol (250 mg, 1 mmol) in pyridine (15 mo) wasadded p-toluenesulfonyl chloride (572 mg, 3 mmol). The light yellowsolution was stirred at r.t. for 17 hours and was then poured into icewater. The mixture was extracted with ether and the combined ethersolution was washed with 5% HCl aq. solution and water. After dryingover anhydrous MgSO₄ the ether solution was concentrated under reducedpressure. The residue was separated by column chromatography to give thecorresponding tosylate.

To a suspension of LiAlH₄ (342 mg, 9 mmol) in THF (30 ml) was added asolution of the tosylate in THF (5 ml). This reaction mixture wasrefluxed for 2 hours. The excess LiAlH₄ was destroyed by addition ofethanol. The mixture was extracted with ether. The combined ethersolution was washed with 2% HCl aq. solution and water. After dryingover anhydrous MgSO₄ the ether solution was concentrated in vacuo.Column chromatography of the residual material afforded 2.11 (245 mg,100% over two steps).

IR (film): 2925, 2865 (s); 1647 (w); 1453, 1378 (s); 731 (s) cm⁻¹.

¹H NMR (500 MHz, CDCl₃): δ5.58 (1H, dt, J=10.2, 3.5); 5.45 (1H, dm,J=10.2); 5.12 (1H, tt, J=7.0, 1.3); 2.02 (1H, m); 1.92 (2H, m); 1.85(1H, tt, J=7.9, 15.4); 1.69 (1H, m); 1.68 (3H, s); 1.59 (3H, s); 1.56(2H, m); 1.46 (1H, m); 1.35 (1H, m); 1.25 (3H, m); 0.93 (3H, s); 0.88(3H, d, J=6.8); 0.78 (3H, d, J=7.1) ppm.

MS (m/z): 234 (1%); 57 (100%).

EXAMPLE 22

Synthesis of Ketone 2.12

To a colourless solution of Hg(OAc)₂ in H₂O (1.25 ml) was added THF(1.25 ml). The reaction mixture became yellow and some precipitateformed. To this mixture was added a solution of 2.11 (212 mg, 0.906mmol) in THF (2.5 ml) and the reaction mixture was stirred at roomtemperature for 2 hours. 3M NaOH (1.09 ml) was added and followed byaddition of 1 M solution of NaBH₄ in 3M NaOH (1.09 ml). This mixture wasstirred for 10 min. Extraction with ether followed by columnchromatography afforded the 25-hydroxy derivative (154 mg) in 67% yield.

IR (film): 3364 (m); 2963, 2867 (s); 1462, 1379 (m); 1153 (m); 732 (m)cm⁻¹.

¹H NMR (500 MHz, CDCl₃): δ5.58 (1H, dt, J=10.3, 3.5); 5.46 (1H, dm,J=10.3); 1.93 (2H, m); 1.70 (1H, m); 1.58 (4H, m); 1.50-1.35 (5H, m);1.26 (2H, m); 1.21 (6H, s); 0.94 (3H, s); 0.88 (3H, d, J=7.0); 0.80 (3H,d, J=7.1) ppm.

MS (m/z) :252 (M.⁺, 1%); 95 (100).

To a solution of the latter (50 mg, 0.20 mmol) in DMF (3 ml) was addedchlorotriethylsilane (90 mg, 0.60 mmol), imidazole (54 mg, 0.80 mmol)and DMAP (10 mg) successively. This solution was stirred at roomtemperature for 20 hours and was then diluted with ether. The ethersolution was washed with 5% HCl aq. solution and water, respectively.After drying over MgSO₄ the solvents were removed in vacuo. The residualmaterial was separated by column chromatography (2% EtOAc in hexane) togive the protected silyl ether (67 mg, 92%).

IR (film): 2958 (s); 1460, 1380 (m); 1235, 1156 §m); 1042 (s); 730 (s)cm⁻¹.

¹H NMR (360 MHz, CDCl₃): δ5.59 (1H, dt, J=10.2, 3.6); 5.46 (1H, dm,J=10.2); 1.92 (2H, m); 1.70 (1H, m); 1.79 (3H, m); 1.45-1.33 (6H, m);1.25 (2H, m); 1.19 (6H, s); 0.94 (9H, t, J=8.0); 0.93 (3H, s); 0.88 (3H,d, J=6.8); 0.80 (3H, d, J=7.3); 0.56 (6H, q, J=8.0) ppm.

MS (m/z): 366 (M.⁺, 1%); 337 (10%); 233 (20%); 173 (30%); 103 (100%).

To a solution of the silyl ether (121 mg, 0.33 mmol) in THF (8 ml) wasadded 9-BBN (0.5M solution in THF, 6.6 ml, 3.3 mmol). This solution wasrefluxed for 30 hours. The organoboranes were oxidized by addingsuccessively EtOH (1 ml), 6N NaOH (0.8 ml), and 30% H₂O₂ (1.6 ml). Thisreaction mixture was heated at 50° C. for 1 hour and was then extractedwith ether. The combined ether solution was washed with 5% HCl aq.solution, water and dried over MgSO₄. After removal of the solvents theresidual material was separated by column chromatography to afford thecyclohexanol (121 mg, 95%).

A mixture of the latter (121 mg, 0.34 mmol) and PDC (480 mg, 1.2 mmol)in CH₂Cl₂ was stirred at room temperature for 20 hours, and wasimmediately filtered through a short column. Purification of the crudeproduct by HPLC (1:20 EtOAc/hexane) furnished cyclohexanone 2.12 (97 mg,80%).

IR (film): 2958 (s); 1722 (s); 1461, 1381 (m); 1282, 1234, 1042 (s); 742(m) cm⁻¹.

¹H NMR (500 MHz, CDCl₃): δ2.28 (1H, d, J=13.2); 2.26 (2H, m); 2.06 (1H,dt, J=13.3, 1.6); 1.90 (1H, m); 1.83 (1H, m); 1.67 (3H, m); 1.18 (6H,s); 0.93 (9H, t, J=8.0); 0.90 (3H, d, J=6.9 Hz); 0.86 (3H, s); 0.79 (3H,d, J=7.2); 0.56 (6H, q, J=8.0) ppm.

MS (m/z): 354 (M.⁺, 10%);353 (5%); 173 (30); 111 (60); 55 (100).

EXAMPLE 23

Synthesis of Alkyne 4.4

A solution of 4.1 (48 g, 0.04 mol), imidazole (6.6 g, 0.68 mol) andt-butyldiphenylchlorosilane (13.2 g, 0.048 mol) in dry DMF (16 ml) isstirred for 36 h at r.t. under nitrogen, then ether (100 ml) is added tothe solution and the organic layer is washed with water (20 ml) threetimes dried over anhydrous MgSO₄ and evaporated to give 15.58 g of 4.2.Purification by column chromatography (hexane:ethylacetate 90:1) gives14.2 g of 4.2 in 100% yield.

Rf: 0.48 (hexane:ethylacetate 5:1).

IR (film): 2932 (m); 1741 (s); 1428 (s); 1199 (s); 1111 (s); 739 (s);702 (s) cm⁻¹.

¹H NMR (500 MHz, CDCl₃): δ7.65 (4H, m); 7.4 (6H, m); 3.82 (1H, dd,J=6.9, 9.7); 3.72 (1H, dd, J=5.8, 9.7); 3.68 (3H, s); 2.72 (1H, sextet,J=6.9); 1.15 (3H, d, J=6.9); 1.03 (9H, s) ppm

To a solution of 4.2 (1.5 g, 4.2 mmol) in dry hexane (9 ml) is furtheradded diisobutyl aluminum hydride (10M/hexane, 4.2 ml, 4.2 mmol)dropwise at −78° C. under nitrogen. Work-up of the reaction with 2Nsolution of potassium sodium tartrate in water under stirring, andsubsequent extraction of the water layer with ether (200 ml), drying ofthe organic layer (MgSO₄, anhydrous) and solvent removal yield 1.32 g ofaldehyde 4.3 contaminated with a small amount of 4.2,

Rf: 0.32 (hexane:ethylacetate 4:1).

¹H NMR (500 MHz, CDCl₃): δ9.76 (1H, d, J=2); 7.65 (4H, m), 7.4 (6H, m),3.87 (2H, m), 2.57 (1H, m); 1.11 (3H, d, J=6.9); 1.03 (9H, s) ppm.

To a suspension of potassium b-butoxide (0.68 g, 6.05 mm) in dry THF (14ml) is added dropwise methyl(diazomethyl)phosphonate (0.59 g, 6.0 mmol)in one minute under nitrogen at −78° C. The resulting red solution isallowed to stir for five minutes at −78° C. and, subsequently, asolution of aldehyde 4.3 (1.78 g, 5.5 mmol) in dry THF (13 ml) is addeddropwise over a one minute period.

The reaction mixture is stirred for 18 h at −78° C. and for 2 h at roomtemperature, and then water (200 ml) is added, the resulting solution isextracted three times with dichloromethane (400 ml) and ether (200 ml).The organic layers are washed with brine, dried over anhydrous MgSO₄,concentrated and purified by column chromatography (hexane:ethyl acetate200:1) to give 1.68 g of 4.4 in 90% yield (from 4.2).

Rf: 0.67 (hexane:ethyl acetate 4:1).

IR (film): 3307 (s); 2959 (m); 2116 (s); 1428 (s); 1112 (s); 702 (s);739 (s) cm⁻¹.

¹H NMR (500 MHz, CDCl₃): δ7.69 (4H, m); 7.4 (6H, m); 3.74 (1H, dd,J=5.7, 9.6); 3.55 (1H, dd, J=7.6, 9.6); 2.66 (1H, ddf, J=2.3, 5.6, 7.6);2.03 (1H, d, J=2); 1.23 (3H, d, J=6.8); 1.07 (9H, s) ppm.

EXAMPLE 24

Synthesis of 4.5

To a well stirred solution of B—Br-9-BBN (1M÷CHCl 8.04 ml, 8.04 mmol) indichloromethane (12 ml) is added dropwise 4.4 (2.16 g; 6.6 mmol) indichloromethane (24 ml) at 0° C. under nitrogen. The reaction mixture isstirred for 4 h at 0° C. Acetic acid (4.26 ml) is then added and themixture is stirred for an additional hour at 0° C., followed by theaddition of 51 ml of 3M NaOH in water and 8.52 ml of 30% hydrogenperoxide. After stirring for 30 min at room temperature (25° C.), theproduct is extracted with hexane three times and the organic layer iswashed with water, aqueous NaHCO₃ and water again and finally dried overMgSO₄ (anh.). The residue obtained after concentration is purified bycolumn chromatography (hexane:ethyl acetate 300:1) to give 2.4 g of 4.5in 90% yield.

Rf: 0.6 (hexane:ethyl acetate 10:1).

IR (film): 2931 (m); 1625 (s); 1427 (s); 1112 (s); 887 (s); 823 (s); 739(s);701 (s) cm⁻¹.

¹H NMR (500 MHz, CDCl₃): δ7.65 (4H, m); 7.42 (6H, m); 5.69 (1H, d,J=1.6); 5.49 (1H, d, J=1.6); 3.71 (1H, dd, J=6.9, 10); 3.56 (1H, dd,J=5.8, 10); 2.61 (1H, overlapped, J=6.9, 6.85, 5.87); 1.09 (3H, d,J=6.85); 1.05 (9H, s) ppm.

EXAMPLE 25

Synthesis of 4.8

To a solution of bromide 4.5 (520 mg, 1.29 mmol) in dry ether (2.5 ml)is added tert-butyllithium (2.6 mmol) rapidly in a portion at −120° C.(excess liquid N₂ in MeOH). To this solution is further added a freshlyprepared solution of CuI (250 mg, 1.29 mmol)/HMPT (484 mg, 2.96 mmol) inether (4.5 ml) at −120° C. The reaction mixture is allowed to warmgradually to −78° C., is further stirred for 1 h and then treated withfreshly distilled BF₃.OEt₂ (310 mg, 2.2 mmol), followed by the dropwiseaddition of 3-methyl-cyclohexenone (116 mg, 1 mmol) in dry ether (2.5ml). The reaction mixture is warmed to −20° C. and left at thistemperature for 10 h. The above solution is poured into aqueous NH₄Cl/6NHCl (4:1 by volume) and extracted with ether (2×50 ml). The combinedextracts are washed with 20% aquous NH₄OH (2×30 ml), 2% aqueous HCl (30ml) and water (30 ml) dried over anhydrous MgSO₄ and evaporated. Theresidue is purified by column chromatography (hexane:ethylacetate 10:1)and HPLC (hexane:ethyl acetate 4:1) to give 174 mg of 4.6 and itsC13-epimer in 40% yield.

Rf: 0.32 (hexane:ethyl acetate 5:1).

To a solution of this mixture (35 mg, 0.08 mmol) in dry THF (1.5 ml) isadded TBAF (1.1 M/THF, 0.3 ml, 0.32 mmol) at room temperature. Afterstirring for 2 h at room temperature the solvent is evaporated and theresidue is purified by column chromatography (H:E 1:1) to give 14.1 mgof 4.7 with its C13-epimer. Careful separation with HPLC (hexane:ethylacetate 6:4; two times) gives pure 4.7 next to its C 13-epimer (1:1).

Rf: 0.27 (hexane:ethyl acetate 1:1).

IR (film): 3386 (s, br); 2932 (m); 2253 (s); 1704 (s); 1590 (m); 1468(s); 1384 (m); 1073 (s) cm⁻¹.

¹H NMR (360 MHz, CDCl₃): δ5.02 (1H, d, J=1.3); 4.94 (1H, s); 3.56 (1H,dd, J=6.3, 10.6); 3.45 (1H, dd, J=7.5, 10.6); 2.58 (1H, AB, d, J=14);2.45 (1H, m); 2.30 (2H, m); 2.22 (1H, AB, d, J=14); 1.82 (2H, m); 1.61(2H, m); 1.10 (3H, s); 1.08 (3H, d, J=6.8) ppm.

EXAMPLE 26 Synthesis of 6.2

To a stirred solution of 6.1 (1 g, 6.50 mmol) and sodium iodide (2.34 g,15.60 mmol) in acetonitrile (12 ml) is added dropwise at 0° C.methyltrichlorosilane (1.84 ml, 15.60 mmol). After 2 hours reflux themixture is cooled and water is added. Extraction with diethylether,followed by washing of the organic phase with aqueous sodiumthiosulfate, water and brine, drying (Na₂SO₄) and concentrating in vacuoyielded the crude iodide which was purified on a silica column(pentane:ethyl acetate 8:2) to give 6.2 (1.58 g; 86.4%).

Rf: 0.21 (pentane:ethyl acetate 85:15).

UV: λ_(max)=254.

IR (film): 3302 (s); 2954 (s); 2866 (s); 1453 (m); 1365 (m); 1262 (m);1183 (m); 1035 (s) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 3.58 (1H, d, J=10.8); 3.43 (1H, d); 3.30(1H, dd, J=9; J=3.5): 2.95 (1H, dd, J=11.5); 2.35-2.10 (2H, m);1.61-1.05 (3H, m); 1.01 (3H, s); 0.98 (3H, s); 0.75 (3H, s) ppm.

EXAMPLE 27

Synthesis of 6.3

To a cooled solution (0° C.) of 6.2 (5 g, 17.73 mmol) in dichloromethane(50 ml) is added dropwise N,N-diisopropylethylamine (DIPEA, 6.96 ml,3.99 mmol) and chloromethyl methyl ether (MOMCI, 1.98 ml, 26.65 mmol).After stirring at room temperature for 3 hours, the mixture is broughtto pH 1-2 and extracted with diethylether (3×). The combined organicfractions are washed with brine and saturated sodium bicarbonate, dried(Na₂SO₄ anh.) and concentrated in vacuo. Purification by columnchromatography (silica; hexane:acetone 95:5) yields 4.95 g (86%) of theMOM diethylether of 6.2. To a stirred solution of this intermediate (870mg, 2.67 mmol) in tetrahydrofuran (6 ml) is added a solution oftetrabutylammonium fluoride (TBAF, 1 M in THF, 21.33 mmol, 21.33 ml).After stirring for 4 hours at room temperature water is added.Extraction with diethylether, drying of the organic phase (MgSO₄) andpurification on a silica column (hexane:acetone 95:5) gives 465 mg pure6.3 (88%).

Rf: 0.76 (hexane:acetone 9:1).

IR (film): 2966 (s); 2877 (s); 1651 (m); 1464 (m); 1369 (m); 1213 (w);1151 (s); 1108 (s); 1049 (s) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 4.76 (2H, dd, J=2.2 Hz); 4.60 (2H, dd,J=6.4 Hz); 3.35 (3H, s); 3.33 (1H, d); 3.25 (1H, d, J=9.3 Hz); 2.43-2.37(2H, m); 1.83-1.75 (1H, m); 1.48-1.38 (1H, m); 0.97 (3H, s); 0.94 (3H,s); 0.92 (3H, s) ppm.

EXAMPLE 28

Synthesis of 6.4 and 6.5

A solution of osmium tetroxide (0.67% in t.butanol, 0.184 mmol, 7 ml) isadded dropwise to a mixture of 6.3 (372 mg, 1.88 mmol) and sodiumperiodate (998 mg, 4.7 mmol) in THF:water 1:1 (4 ml). After stirring for30 hours at rom temperature, a saturated sodium thiosulfate solution inwater (1 ml) is added and the resulting mixture is extracted withdichloromethane. Purification by column chromatography (silica;hexane:ethyl acetate 93:7) yields 244 mg (65%) of the ketone. A solutionof this ketone (244 mg, 1.22 mmol) in tetrahydrofuran (1 ml) is addeddropwise to a suspension of lithium aluminum hydride (47 mg, 1.22 mmol)in tetrahydrofuran (2 ml). After stirring at room temperature for 1hour, sodium sulfate decahydrate is added and the resulting mixture isstirred for an additional 2 hours and subsequently filtered to removethe metal salts. The filtrate is concentrated in vacuo and purified bycolumn chromatography (silica; hexane:ethyl acetate 85:15) to give 222mg (90%) of a mixture of diastereomers 6.4 and 6.5.

Rf: 0.16 (hexane:ethyl acetate 8:2).

IR (film): 3426 (s, br); 2958 (s); 2876 (s); 1467 (m); 1369 (m); 1216(m); 1151 (s); 1108 (s); 1046 (s) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 4.63 (2H:2, s); 4.59 (2H:2, s); 3.79 (1H:2,dd); 4.01 (1H:2, dd); 3.41 (1H:2, d); 3.30 (1H:2, d); 3.29 (1H:2, d);3.25 (1H:2, d); 3.39 (3H:2, s); 3.36 (3H:2, s); 2.20-1.40 (4H, m);1.00-0.85 (9H, 3×s) ppm.

EXAMPLE 29

Synthesis of 6.6

To a solution of 6.3 (1.552 g, 7.84 mmol) in tetrahydrofuran (35 ml) isadded a solution of 9 borabicyclo[3.3.1]nonane (0.5 M intetrahydrofuran, 15.7 ml, 7.85 mmol) and the resulting solution isstirred for 5 hours at 55° C. After cooling to room temperature, ethanol(4.71 ml) and a 6 M aqueous sodium hydroxide solution (1.57 ml, 9.42mmol) are added, followed by dropwise addition at 0° C. of a 35% aqueoussolution of hydrogen peroxide (3.68 ml). Stirring for 1 hour at refluxtemperature, subsequent extraction of the water layer with diethylether,drying of the organic phase (Na₂SO₄ anh.) and solvent removal yields 2.8g of a crude oil. Purification by column chromatography (hexane:acetone8:2) and HPLC (silicagel; hexane:ethyl acetate 75:25) gives 6.6 (617 mg,36%) next to the undesired epimer (864 mg, 51%)

Rf:0.25 (dichloromethane:methanol 9:1).

IR (film): 3418 (s, br); 2946 (s); 2875 (s); 1464 (s); 1368 (m); 1215(m); 1150 (s); 1108 (s); 1047 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.61 (2H, 2×d, J=6.5); 3.72 (1H, dd, J=5.6,10.2); 3.52 (1H, dd, J=10.2, 9.3); 3.35 (3H, s); 3.34 (1H, d, J=9.15);3.25 (1H, d); 2.10 (1H, ddd), 1.88 (2H, m); 1.65 (1H, m); 1.43 (1H, m);1.35 (1H, m); 0.91 (3H, s); 0.90 (3H, s); 0.83 (3H, s) ppm.

EXAMPLE 30

Synthesis of 6.7

To a mixture of triphenylphosphine (1.46 g, 5.56 mmol), imidazole (378mg, 5.56 mmol) and 6.6 (600 mg, 2.78 mmol) in diethylether:acetonitrile3:1 (12 ml) is added at 0° C., portionwise, iodine (1.41 g, 5.56 mmol)and the resulting mixture is stirred in the dark for 3 hours at roomtemperature. Extraction with diethylether:hexane 1:1, washing of thecollected organic fractions with brine, drying (Na₂SO₄ anh.) and solventremoval gives a pale yellow oil which is purified by columnchromatography (silicagel; hexane:ethyl acetate 95:5) to yield 825 mg(91%) of the iodide. A solution of this ether (460 mg, 1.41 mmol) inmethanol:tetrahydrofuran 3:1 (70 ml) is stirred in the presence ofAmberlyst-15 for 72 hr at room temperature in the dark. Afterwards theAmberlyst-15 is filtered off and the filtrate is concentrated in vacuoand purified by column chromatography (silicagel; hexane:ethyl acetate85:15 to 70:30) to give 6.7 (342 mg, 86%).

Rf: 0.20 (hexane:ethyl acetate 8:2).

IR (film): 3380 (s); 2963 (s); 2872 (m); 1452 (m); 1368 (m); 1264 (m);1183 (m); 1028 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 3.41 (2H, s br); 3.30 (1H, dd, J=9, 3.6);2.98 (1H, dd, J=11.5); 2.35-2.27 (1H, m); 2.13-2.05 (1H, m); 1.81-1.73(1H, m); 1.44-1.30 (3H, m); 0.89 (3H, s); 0.88 (3H, s); 0.71 (3H, s)ppm.

EXAMPLE 31

Synthesis of 6.8

To a suspension of copper(I) iodide (676 mg, 3.55 mmol) and zinc dust(928 mg, 14.18 mmol) in ethanol:water 7:3 (10 ml) are added methyl vinylketone (380 μl, 4.62 mmol) and 2 (1 g, 3.55 mmol). The reaction mixtureis sonicated during 1 hour under an argon atmosphere followed byaddition of more copper(I) iodide (338 mg, 1.775 mmol) and zinc dust(464 mg, 7.09 mmol). After another 35 minutes sonication, the mixture isfiltered through celite and the copper- and zinc salts are washed withethyl acetate. The filtrate is extracted with ethyl acetate, dried(anhydric magnesiumsulfate) and concentrated. Purification on a silicacolumn (pentane:ethyl acetate 7:3) gives 510 mg 6.8 (64%).

Rf: 0.29 (pentane:ethyl acetate 85:15).

IR (film): 3420 (s); 2946 (s); 2869 (s); 1714 (s); 1454 (s); 1367 (s);1231 (m); 1163 (s); 1024 (s) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 3.58 (1H, d, J_(6.6′)=10.7); 3.45 (1H, d);2.50-2.35 (2H, m); 2.14 (3H, s); 1.96-1.84 (1H, m); 1.80-1.13 (8H, m);0.98 (3H, s); 0.88 (3H, s); 0.70 (3H, s) ppm.

EXAMPLE 32

Synthesis of 6.10

To a solution of methyllithium (1.5 M in diethylether 11.6 ml, 17.34mmol) is added dropwise a solution of 6.8 (490 mg, 2.17 mmol) indiethylether (5 ml) at −78° C. After stirring under an argon atmospherefor 2 hours, saturated ammonium chloride is added and the resultingmixture is extracted with diethylether, the organic phase dried (MgSO₄)and the solvent removed. Purification of the crude by columnchromatography (silica; pentane:ethyl acetate 8:2) yields 455 mg of thewhite crystalline diol (85%). A solution of this diol (200 mg, 0.826mmol) in dichloromethane (0.8 ml) is added to a mixture of dipyridinechromium(VI)oxide (1.066 g, 4.13 mmol) and dichloromethane. The mixtureis stirred at room temperature; after two hours diethylether (5 ml) andcelite are added. Filtration through silicagel-celite, washing withdiethylether and solvent removal gives a residue which is purified on asilica column (pentane:ethyl acetate 8:2) and by HPLC (pentane:ethylacetate 75:25). 70 mg of pure 6.10 is obtained (35%).

Rf: 0.24 (pentane:ethyl acetate 85:15).

IR (film): 3480 (s); 2964 (s); 1715 (s); 1470 (m); 1372 (m) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 9.64 (1H, s); 2.39-2.28 (1H, m); 2.04-1.92(1H, m); 1.83-1.72 (1H, m); 1.53-1.23 (8H, m); 1.21 (6H, s); 1.03 (3H,s); 0.96 (3H, s); 0.75 (3H, s) ppm.

EXAMPLE 33

Synthesis of 6.12

A solution of n.butyllithium (2.5 M in hexane, 467 μl, 1.17 mmol) isadded dropwise to a solution of (methoxymethyl)-triphenylphosphoniumchloride (400 mg, 1.17 mmol) in tetrahydrofuran (4 ml) at −78° C. Theresulting mixture is stirred for 20 minutes and subsequently a solutionof 6.10 (70 mg, 291 μmol) in tetrahydrofuran (700 μl) is added dropwise.After stirring for 10 minutes, at −78° C., the mixture is allowed towarm to room temperature and stirred for an additional 21 hours.Addition of water, extraction with diethylether, drying (Na₂SO₄) andconcentrating gives the crude enolether. To a solution of the enolether(65 mg, 243 μmol) in tetrahydrofuran (700 μl) is added a hydrochloricacid solution (2 M in THF, 66 μl). After 30 minutes the mixture isextracted with diethylether, the organic portions are washed withsaturated sodium bicarbonate and brine and dried (Na₂SO₄). After removalof the solvent with a rotary evaporator the remaining oil is purified bycolumn chromatography (silica; pentane:ethyl acetate 8:2); 25 mg pure6.12 is obtained (45%).

Rf: 0.17 (pentane:ethyl acetate 8:2).

IR (film): 3440 (s); 2928 (m); 1715 (m); 1470 (w); 1380 (w) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 9.84 (1H, dd, J=4.1, 2.4); 2.31 (2H, dd);1.99-1.90 (1H, m); 1.87-1.23 (1OH, m); 1.22 (6H, s); 0.98 (3H, s); 0.82(3H, s); 0.68 (3H, s) ppm.

EXAMPLE 34

Synthesis of 6.9

To a suspension of copper(I)iodide (676 mg, 3.55 mmol) and zinc dust(928 mg, 14.18 mmol) in ethanol:water 7:3 (10 ml) are added ethyl vinylketone (458 μl, 4.61 mmol) and 6.2 (1 g, 3.55 mmol). The reactionmixture is sonicated during 1 hour under an argon atmosphere followed byaddition of more copper(I)iodide (338 mg, 1.775 mmol) and zinc dust (464mg, 7.09 mmol). After another 35 minutes of sonication, the mixture isfiltered through celite and the copper and zinc salts are washed withdiethylether in the sonicator. After drying on sodium sulfate, thefiltrate is concentrated and purified on a silica column (pentane:ethylacetate 85:15) to give 433 mg 6.9 (51%).

Rf: 0.29 (pentane:ethyl acetate 85:15).

IR (film): 3473 (s, br); 2940 (s); 2871 (s); 1712 (s); 1460 (s); 1376(s); 1113 (s); 1028 (s) cm⁻¹.

¹H NMR: (200 MHz, CDCl₃): δ: 3.52 (2H, 2×d, J=10.6); 2.42 (2H, q,J=7.3); 1.98-1.14 (11H, m); 1.06 (3H, t, J=7.3); 0.98 (3H, s); 0.89 (3H,s); 0.70 (3H, s) ppm.

EXAMPLE 35

Synthesis of 6.11

To a solution of ethyl iodide (305 μl , 3.75 mmol) in diethylether (3.75ml) is added tert.butyllithium (3.21 ml of a 2.34 M sol. in pentane, 7.5mmol) at −78° C. and the resulting solution is stirred for 1 hour.Subsequently a solution of 6.9 (300 mg, 1.25 mmol) in dry diethylether(3 ml) is added dropwise. The mixture is stirred for 2 hours at −78° C.under an argon atmosphere and then brought to room temperature.Saturated ammonium chloride is added and the resulting mixture isextracted with diethylether and dichloromethane. The organic phase isdried (MgSO₄), filtered, concentrated and purified on a silica column(pentane:ethyl acetate 8:2) to yield 251 mg (74%) of the diol.

To a mixture of 4-methylmorpholine N-oxide (158 mg, 1.35 mmol),activated powdered molecular sieves 4 Å (450 mg) and the diol (243 mg,0.9 mmol) in dichloromethane (1.8 ml) is added at 0° C.tetra(n.propyl)ammonium perruthenate (15.8 mg, 45 μmol) in portions.After 2 hours stirring at room temperature, the reaction mixture isfiltered through silicagel, washed with dichloromethane and concentratedin vacuo. Purification by column chromatography (silica; pentane:ethylacetate 85:15) gives 193 mg 6.11 (80%).

Rf: 0.20 (pentane:ethyl acetate 9:1).

IR (film): 3436 (s, br); 2965 (s); 2938 (s); 2877 (s); 1721 (s); 1460(s); 1370 (m); 1266 (m); 1186 (m) cm⁻¹.

¹H NMR: (200 MHz, CDCl₃): δ: 9.65 (1H, s); 2.48-2.25 (1H, m); 2.09-1.90(1H, m); 1.88-1.70 (1H, m); 1.65-1.20 (13H, m); 1.02 (3H, s); 0.97 (3H,s); 0.87 (6H, t, J=7.4 Hz); 0.76 (3H, s) ppm.

EXAMPLE 36

Synthesis of 6.13

To a solution of (methoxymethyl)triphenylphosphonium chloride (330 mg,963 μmol) in diethylether (2.5 ml) is added n.butyllithium (2.5 M sol.in hexane, 347 μl, 866 μmol) at 0° C. After stirring for 10 minutes thered suspension is brought to room temperature, stirred 10 minutes andsubsequently cooled again to −30° C. A solution of 6.11 (86 mg, 321lmol) in diethylether (860 μl) is added dropwise and after ½ hour thecooling bath is removed and the mixture stirred at room temperature for18 hours. Addition of water, followed by extraction with diethylether,drying (Na₂SO₄) and concentration in vacuo yields 200 mg of crudevinylether. After filtering through silicagel and evaporation of thesolvent, the filtrate is diluted in tetrahydrofuran (1 ml) and treatedwith aqueous hydrochloric acid (2N sol. in tetrahydrofuran). After 30min water is added and the mixture is extracted with diethylether, dried(Na₂SO₄) and filtered. The filtrate is concentrated in vacuo andpurified on a silica column (pentane:ethyl acetate 9:1) and by HPLC(hexane:ethyl acetate 8:2) to give 30 mg (33%) of 6.13.

Rf: 0.26 (pentane:ethyl acetate 9:1).

IR (film): 3455 (m, br); 2965 (s); 2938 (s); 2876 (m); 1720 (s); 1460(m); 1144 (m) cm⁻¹.

¹H NMR: (200 MHz, CDCl₃): δ: 9.8 (1H, dd, J=3, 3.5 Hz); 2.15 (2H, 2×d,J<1, 3, 3.5 Hz); 2.05-1.52 (2H, m); 1.52-1.4 (4H, q, J=7.5 Hz); 1.4-1.0(6H, m); 0.99 (3H, s); 0.86 (6H, t, J=7.5 Hz); 0.81 (3H, s); 0.69 (3H,s) ppm.

EXAMPLE 37

Synthesis of 6.17(α+β) (R=Me)

A mixture of 6.4 and 6.5 (730 mg, 3.61 mmol)e potassium hydroxide(powdered, 400 mg, 7.22 mmol) and 1-chloro-3-methyl-2-butene (610 μl,5.42 mmol) in toluene (8 ml) is sonicated during 30 minutes. Afteraddition of a trace of 18-Crown-6 and more potassium hydroxide (200 mg,3.61 mmol) the mixture is sonicated for an additional hour. Subsequentlythe mixture is filtered through a short pad of silicagel, theprecipitate is washed with diethylether, the filtrate concentrated andpurified by column chromatography (hexane:ethyl acetate 95:5→8:2) whichyields 286 mg 6.17(α+β) (R=Me) (29%) and 450 mg of unreacted material.

Rf: 0.63 (hexane:ethyl acetate 8:2).

IR (film): 3015 (s); 1617 (m); 1420 (m); 1215 (s), 1015 (m); 923 (s)cm⁻¹.

¹H NMR: in accordance with structures of both epimers.

EXAMPLE 38

Synthesis of 6.18(α+β) (R=H)

A mixture of freshly powdered potassium hydroxide (700 mg, 12.5 mmol),18-Crown-6 ether (50 mg, 193 mmol), 6.4 and 6.5 (700 mg, 3.47 mmol) andallyl bromide (644 μl, 7.62 mmol) in tetrahydrofuran (7 ml) is stirredfor 48 hours at room temperature. The mixture is filtered throughsilicagel and the filtrate concentrated in vacuo. Purification by columnchromatography (silica; hexane:ethyl acetate 95:5) yields 630 mg6.18(α+β) (R=H) (75%).

Rf: 0.67 (hexane:ethyl acetate 8:2).

IR (film): 3079 (w); 2956 (m); 2875 (); 1 64 6 (w); 1465 (m); 1371 (m);1150(s); 1106 (s); 1049 (s); 918(s) cm⁻¹.

¹H NMR: in accordance with structures of both epimers.

EXAMPLE 39

Synthesis of 6.19α (R=Me) and 6.19β (R=Me)

To a suspension of mercuric acetate (432 mg, 1.36 mmol) in water (1.35ml) and tetrahydrofuran (1.35 ml) is added a solution of 6.17(α+β)(R=Me) (307 mg, 1.138 mmol) in tetrahydrofuran (2.7 ml); after a fewminutes the color of the mixture becomes pale yellow. The mixture isstirred for 1 hour at room temperature and subsequently a 3M aqueoussodium hydroxide solution (1.35 ml) is added, immediately followed byaddition of a sodium borohydride solution (1M in 3M sodium hydroxide,0.68 ml). This yields a dark grey suspension which is filtered over ashort pad of silicagel. The concentrated filtrate is purified by columnchromatography (silica; hexane:ethyl acetate 8:2) to give 308 mg 19(α+β)(R=Me) (94%) of the tertiary alcohol. To a solution of this (288 mg, 1.0mmol) in methanol:tetrahydrofuran 2:1 (90 ml) is added Amberlyst 15 (32g). The resulting mixture is stirred for 55 h at room temperature andsubsequently filtered through silicagel. The filtrate is concentrated invacuo and purified by column chromatography (silica; hexane:ethylacetate 6:4) to yield the diol (240 mg; 97%). To a mixture of4-methylmorpholine N-oxide (NMMO, 157 mg, 1.348 mmol), activatedpowdered molecular sieves 4 Å (449 mg) and the diol (230 mg, 0.94 mmol)in dry dichloromethane (3 ml) is added portionwise at −10° C., solidtetra (n.propyl)ammonium perruthenate (TPAP, 15.8 mg, 45 lmol). Afterstirring for 1½ h at room temperature the mixture is filtered throughcelite and the residue washed with ethyl acetate. The dark coloredfiltrate is concentrated at the rotavapor and purified by columnchromatography (silica; hexane:ethyl acetate 7:3). The two C-20diastereomers 6.19α (R=Me; 90 mg, 40%) and 6.19p (R=Me; 60 mg, 26%) areseparated by HPLC (hexane:acetone 92:8) and the relative configurationof both is established by NOE-experiments.

19α: Rf: 0.36 (hexane:acetone 75:25).

IR (film): 3444 (s, br); 2969 (s); 2875 (s); 1718 (s); 1466 (s); 1367(s); 1161 (s); 1087 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 9.65 (1H, s); 3.76-3.71 (1H, ddd);3.55-3.49 (2H, m); 2.38-2.32 (1H, ddd); 2.12-2.04 (1H, m); 1.75-1.72(2H, t); 1.72-1.68 (1H, m); 1.45-1.38 (1H, ddd); 1.23 (6H, s); 1.01 (3H,s); 0.99 (3H, s); 0.95 (3H, s) ppm.

19β: Rf: 0.41 (hexane:acetone 75:25).

IR (film): 3446 (s, br); 2964 (s); 2872 (s); 1718 (s); 1466 (s); 1384(s); 1367 (s); 1154 (s); 1098 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 9.61 (1H, s); 3.79-3.73 (1H, ddd, J=5.6,5.7 Hz); 3.60-3.54 (1H, ddd); 3.50-3.45 (1H, t, J=7.3 Hz); 2.18-2.03(2H, m); 1.77-1.74 (2H, t); 1.65-1.57 (2H, m); 1.52-1.45 (1H, m); 1.23(6H, s); 1.09 (3H, s); 1.00 (3H, s); 0.90 (3H, s) ppm.

EXAMPLE 40

Synthesis of 6.20α (R=Et) and 6.20β (R=Et)

A solution of 6.18α,β (R=H) (600 mg, 2.48 mmol) and 9-borabicyclo[3.3.1]nonane (0.5 M in THF, 19.8 ml, 9.92 mmol) in tetrahydrofuran isstirred for 5 hours at 55° C. The mixture is brought to roomtemperature, ethanol (5.26 ml) and a 6M aqueous sodium hydroxidesolution (1.75 ml, 9.92 mmol) are added and the mixture is subsequentlycooled to 0° C. A 35% aqueous hydrogen peroxide solution (4.2 ml) isadded slowly, followed by refluxing for 1 hour. Extraction withdiethylether, drying (MgSO₄) and solvent evaporation yields a residuewhich is purified by column chromatography (silica; hexane:ethyl acetate6:4) to give 615 mg of the alcohol (95%).

A mixture of this alcohol (220 mg, 0.846 mmol) and pyridinium dichromate(1.43 g, 5.08 mmol) in N,N-dimethylformamide (6ml) is stirred for 12hours at 40° C. Water is added and the mixture extracted withdiethylether. Drying of the organic pnase (MgSO₄) and concentration invacuo yields a yellow oil which is diluted in diethylether. The solutionis cooled at 0° C. and a solution of diazomethane in diethylether isadded dropwise till complete methylation is observed by thin layerchromatography. Subsequently an equal volume of hexane is added and theorganic phase is washed with water, dried (MgSO₄) and concentrated invacuo. Purification by column chromatography (silica; hexane/ethylacetate 8:2) and HPLC (hexane:ethyl acetate 9:1) yields 85 mg (35%) ofthe methylester.

A solution of this ester (90 mg, 0.31 mmol) in diethylether is added toethylmagnesium iodide (1.3 mmol). The resulting mixture is stirred for 2hours at room temperature and subsequently quenched with saturatedammonium chloride. Extraction with diethylether, drying of the organicfraction (MgSO₄) solvent removal and purification on silica(hexane:ethyl acetate 7:3) gives 90 mg (92%) of the tertiary alcohol.

To a solution of this alcohol (90 mg, 0.285 mmol) inmethanol:tetrahydrofuran 3:1 (20 ml) is added Amberlyst 15 (7 g). After72 hours stirring at room temperature the Amberlyst is filtered off, thefiltrate is concentrated in vacuo and purified on a silica column(hexane:ethyl acetate 6:4) to yield 65 mg (84 %) of the diol.

To a solution of this diol (50 mg, 0.184 mmol) and triethylamine (211μl, 1.84 mmol) in dimethyl sulfoxide:dichloromethane 1:1 (2 ml) is addedportionwise sulfur trioxide pyridine complex (179 mg, 1.104 mmol). After2 hours stirring, under nitrogen, at room temperature the mixture isfiltered through silicagel and the filtrate, after solvent removal,purified by column chromatography (silica; hexane:acetone 9:1). HPLC(hexane:acetone 92:8) separation gives the two epimeric alcohols 6.20α(R=Et, 13 mg, 26%) and 6.20β (R=Et, 20 mg, 40%). The relativestereochemistry is established by NOE experiments.

6.20α: Rf: 0.32 (hexane:ethyl acetate 8:2).

IR (film): 3519 (s, br); 2966 (s); 2878 (s); 2728 (w); 1718 (s); 1462(s); 1371 (m); 1264 (w); 1138 (s); 1089 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 9.65 (1H, s); 3.71 (1H, m); 3.50 (2H, m);2.39-2.31 (1H, m); 2.12-2.04 (1H, m); 1.75-1.67 (3H, m); 1.55-1.38 (6H,m); 1.01 (3H, s); 0.99 (3H, s); 0.95 (3H, s); 0.86 (6H, t) ppm.

6.20β: Rf: 0.35 (hexane:ethyl acetate 8:2).

IR (film): 3516 (s, br); 2962 (s); 2877 (s); 2716 (m); 1721 (s); 1463(s); 1368 (m); 1139 (s); 1100 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 9.61 (1H, s); 3.75 (1H, m); 3.54 (1H, m);3.47 (1H, t); 2.17-2.03 (2H, m); 1.72 (2H, t); 1.65-1.42 (7H, m); 1.10(3H, s); 1.00 (3H, s); 0.90 (3H, s); 0.86 (6H, t) ppm.

EXAMPLE 41

Synthesis of 621α (R=Me)

A solution of n.butyllithium (2.5 M in hexanes, 195 μl, 0.486 mmol) isadded dropwise at −10° C. to a suspension of (methoxymethyl) triphenylphosphonium chloride (233 mg, 0.680 mmol) in diethylether (1.8 ml).After 20 minutes the resulting red suspension is brought to roomtemperature, stirred for 10 minutes and then cooled again to −30° C. Asolution of 6.19α (R=Me) (47 mg, 194 μmol) in diethylether (0.5 ml) isadded dropwise, after stirring for ½ h at −30° C. the mixture is broughtto room temperature and stirred for 15 hours. Work-up by filtrationthrough silicagel, washing of the residue with diethylether andconcentration of the filtrate yields 64 mg of a pale yellow oil which isdiluted in tetrahydrofuran (1 ml). A solution of hydrochloric acid (2 Nin tetrahydrofuran, 120 μl) is added and the resulting solution isstirred for 2 h at room temperature. Filtration through silicagel,concentration of the filtrate and purification on HPLC (hexane:acetone9:1) give 6.21α (R=Me; 24 mg; 48%).

Rf: 0.21 (hexane:ethyl acetate 85:15).

¹H NMR: (360 MHz, CDCl₃): δ: 9.82 (1H, dd, J=2.2, 4 Hz); 3.79-3.72 (1H,dt, J=5.7, 9.5); 3.64-3.56 (1H, dd, J=5.6 Hz); 3.61-3.54 (1H, dt);2.42-2.37 (1H, dd, J=2.2, 14.5 Hz); 2.33-2.27 (1H, dd, J=4, 14.5 Hz);2.19-2.09 (1H, m); 1.99-1.89 (1H, m); 1.75 (2H, t); 1.66-1.54 (3H, m);1.23 (6H, s); 1.00 (3H, s); 0.90 (3H, s); 0.83 (3H, s) ppm.

EXAMPLE 42

Synthesis of 6.21β (R=Me)

As described for 6.21α (R=Me; yield 36%).

Rf: 0.15 (hexane:ethyl acetate 85:15).

IR (film): 3452 (s, br); 2968 (s); 2877 (m); 1720 (s); 1468 (m); 1366(m); 1155 (m); 1094 (s) cm⁻¹.

¹H NMR: (200 MHz, CDCl₃): δ: 9.84 (1H, dd, J=3.6 Hz); 3.81-3.42 (3H, m);2.38-2.01 (4H, m); 1.80-1.55 (5H, m); 1.22 (6H, 2xs); 1.10 (3H, s); 0.88(6H, 2×s) ppm.

EXAMPLE 43

Synthesis of 6.22α (R=Et)

A solution of n.butyllithium (2.5 M in hexane, 57 μl, 0.142 mmol) isadded dropwise at −10° C. to a suspension of (methoxymethyl) triphenylphosphoniumchloride (56 mg, 0.163 mmol) in diethylether (0.8 ml). After10 minutes the resulting red suspension is brought to room temperaturestirred for 10 minutes and then cooled again to −30° C. A solution of6.20a α (11 mg, 40.7 μmol) in diethylether (0.2 ml) is added dropwise,after ½ hour at −30° C. the mixture is brought to room temperature andstirred for 15 hours. Work-up by filtration through silicagel, washingof the residue with diethylether and concentration of the filtrateyields 35 mg of a pale yellow oil which is diluted in tetrahydrofuran (1ml). A solution of hydrochloric acid (2N in tetrahydrofuran, 150 μl) isadded and the resulting solution is stirred for 2 h at room temperature.Filtration through silicagel, concentration of the filtrate andpurification on HPLC (hexane:acetone 9:1) gives 3 mg (26%) of 6.22α.

Rf: 0.27 (hexane:ethyl acetate 8:2).

¹H NMR: (500 MHz, CDCl₃): δ: 9.81 (1H, t); 3.72 (1H, m); 3.52 (2H, m);2.40 (1H, dd); 2.31 (1H, dd); 2.18-2.08 (1H, m); 1.99-1.90 (1H, m);1.75-1.38 (9H, m); 1.01 (3H, s); 0.91 (3H, s); 0.87 (3H, s); 0.85 (6H,t) ppm.

EXAMPLE 44

Synthesis of 6.22β (R=Et)

As described for 6.22α (R=Et); yield 36%.

Rf: 0.29 (hexane:ethyl acetate 8:2).

IR (film): 3513 (s, br); 2963 (s); 2879 (m); 2732 (w); 1720 (s); 1463(m); 1387 (m); 1094 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 9.80 (1H, dd); 3.72 (1H, m); 3.49 (2H, m);2.30 (1H, dd, J=14.3, 3.5 Hz); 2.21 (1H, dd, J=2.6 Hz); 2.15-2.08 (1H,m); 1.80-1.42 (10H, m); 1.10 (3H, s); 0.89 (3H, s); 0.87 (3H, s); 0.86(6H, t) ppm.

EXAMPLE 45

Synthesis of 6.16

Starting from 6.7 as described for the synthesis of 6.13 starting from6.3.

6.14: Rf: 0.26 (hexane:ethyl acetate 8:2).

¹H NMR: (360 MHz, CDCl₃): δ: 3.42 (2H, s, br); 2.42 (4H, m); 1.90-1.24(9H, m); 1.05 (4H, t, J=7.3 Hz); 0.89 (3H, s); 0.80 (3H, s); 0.67 (3H,s) ppm.

6.15: Rf: 0.21 (hexane:ethyl acetate 8:2).

¹H NMR: (500 MHz, CDCl₃): δ: 9.65 (1H, s); 2.00 (2H, m); 1.65 (2H, m);1.45 (4H, q); 1.40-1.05 (8H, m); 1.01 (3H, s); 0.93 (3H, s); 0.85 (6H,t); 0.71 (3H, s) ppm.

6.16: Rf: 0.26 (hexane:ethyl acetate 8:2).

IR (film): 3426 (s, br); 2935 (s); 1714 (m); 1650 (s); 1390 (m); 1112(m) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 9.86 (1H, t, J=3.1 Hz); 2.29 (1H, dd,J=14.5 Hz); 2.24 (1H, dd); 1.91 (1H, m); 1.75 (1H, m); 1.66 (1H, m);1.59 (1H, m); 1.45 (4H, q, J=7.6 Hz); 1.42-1.07 (8H, m); 1.05 (3H, s);0.86 (6H, t); 0.80 (3H, s); 0.69 (3H, s) ppm

EXAMPLE 46

Synthesis of 6.24

To a solution of 6.2 (1.9 g, 6.7 mmol) in CH₂Cl₂ (120 ml) at 0° C.,DIPEA (20 eq, 20 ml) is added. After stirring for 40 min at 0° C. MEMCI(8 eq, 6 ml) is added and stirring is continued for 2 h. The mixture ispoured in a water-ether mixture. The organic phase is dried (MgSO₄).After filtration and evaporation the residue is purified by columnchromatography (silicagel, diethylether:hexane 1:3) giving 6.23 (2.02 g,80%).

Rf: 0.49 (diethylether:hexane 1:1).

IR (film): 3480, 3308, 1782, 1150, 1085cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.688 and 4.671 (2×1H, J=6.7); 3.68 and3.56 (2×2H, 2×m); 3.47 (1H, d, J=9.3), 3.39 (3H, s); 3.297 (1H, dd,J=3.3, 9.0); 3.281 (1H, d, J=9.4); 2.94 (1H, dd, J=9.0, 11.7); 2.28 (2H,dtd, J=3, 10, 12); 2.24 (1H, m); 0.99 (3H, s); 0.96 (3H, s); 0.72 (3H,s) ppm.

To a solution of 6.23 (1 g, 2.7 mmol) in DMF (160 ml), sodium nitrite(400 mg, 2 eq) and a catalytic amount of urea is added. After stirringfor 2 days at r.t. the solution is poured in a ether-ice mixture. Theether phase is dried (MgSO₄). After filtration and evaporation theresidue is purified by column chromatography (silicageldiethylether:hexane 1:6→1:3) giving the nitro compound (350 mg; 45%)with Rf=0.36 (diethylether:hexane 1:1).

To a solution of the nitro compound (345 mg, 1.2 mmol) in anhydrous MeOH(25 ml) NaOMe (98 mg, 1.3 eq), is added. After stirring for 30 min, thesolution is cooled to −78° C. and a ozone flow (20 mmol/h) is passedthrough until the colour is deep blue (30 min), then the solution isflushed with nitrogen for 30 min at −78° C. followed by addingdimethylsulphide (3.5 ml).

The mixture is warmed to r.t. and after solvent evaporation anether-brine mixture is added. The ether phase is dried (MgSO₄). Afterfiltration, solvent evaporation and column chromatography (silicagel,nitromethane:benzene 1:14) gives 6.24 (215 mg, 70%).

Rf: 0.14 (nitromethane:benzene 1:14).

IR (film): 1717 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 9.76 (1H, d, J=2.24); 4.692 and 4.675(2×1H, J=6.7); 3.69 and 3.56 (4H); 3.49 (1H, d, J=9.4); 3.39 (3H, s);3.32 (1H, d, J=9.4); 2.69 (1H, td, J=2.2, 9.1); 2.11 (1H, m); 1.20 (3H,s); 1.02 (3H, s); 0.91 (3H, s) ppm.

EXAMPLE 47

Synthesis of 6.25

To a solution of n.butyllithium (500 μl, 2.4 M, 1.5 eq, in hexane) inTHF (6 ml) at −78° C., under argon atmosphere, diisopropylamine (1.5 eq,168 μl) is added. After stirring for 20 min at −78° C.triethyl-4-phosphonocrotonate (1.5 eq, 333 mg, 90%, 300 μl) is addeddropwise. After stirring for 2 h at −78° C., a solution of 6.24 (215 mg,833 μmol, 1 eq) in THF (5 ml) is added dropwise and stirring iscontinued for 2 h at −78° C. The mixture is then slowly warmed up tor.t. and is poured in an ether-brine mixture. The ether phase is dried(MgSO₄).

Filtration, evaporation and column chromatography (diethylether:hexane1:4) gives the dienic ester (267 mg, 91% with Rf=0.39(diethylether:hexane 1:1). To a solution of this product (267 mg, 754μmol) in EtOAc (10 ml), a catalytic amount of palladium on carbon (10%)is added, after which the mixture is hydrogenated for 3 h (4 atm).Filtration over celite, addition of Et₃N (200 μl), evaporation andcolumn chromatography (diethylether:hexane 1:14→1:6) gives the saturatedproduct (215 mg, 80%, with Rf=0.53 (diethylether:hexane 1:1)).

To a solution of this (60 mg, 169 μmol) in CH₂Cl₂ (1200 μl), at −78° C.,a solution of dimethylborobromide (±10 eq, 1 ml, 1.5 M inCH₂Cl₂:ClCH₂CH₂Cl 2:1) is added. After stirring for 1 h at −78° C., themixture is transferred to a vigorously stirred mixture of THF (8 ml) andsaturated NaHCO₃ solution (4 ml). The reaction flask is washed withdichloromethane (2×2 ml:) followed by addition of ether and brine. Theorganic phase is dried (MgSO₄). After filtration and evaporation andcolumn chromatography (diethylether:hexane 1:3) 6.25 (42 mg, 93%) isobtained.

Rf: 0.38 (diethylether:hexane 1:1).

IR (film):1717 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.12 (2H, q, J=7.13); 3.57 (13H, br, d,J=10), 3.45 (1H, br, d, J=10); 2.29 (2H, m); 1.87 (1H, m); 1.73 (1H, qd,J=2, 10); 1.25 (3H, t, J=7.13); 0.98 (3H, s); 0.89 (3H, s); 0.71 (3H, s)ppm.

EXAMPLE 48

Synthesis of 6.26

To a solution of 6.25 (140 mg, 520 μmol) in CH₂Cl₂-DMSO (2.5 ml:5 ml),at −15° C., a solution of Et₃N (3 eq, 220 μl) and sulphurtrioxide-pyridine complex (25 eq, 205 mg) in CH₂Cl₂-DMSO (1 ml:2 ml) isadded dropwise. After stirring for 3 h between −10° C. and 4° C, themixture is poured in an ether-brine. The organic phase is dried (MgSO₄).Filtration, evaporation, and column chromatography (silicagel,diethylether:hexane 1:9) gives the aldehyde (100 mg, 72% with Rf=0.60(diethylether:hexane 1:1)). This aldehyde is transformed into 6.26 asdescribed for 6.12 from 6.10 (yield 57%).

Rf: 0.50 (Et₂O:hexane 1:1).

¹H NMR: (500 MHz, CDCl₃): δ: 9.83 (1H, dd, J=2.48, 4.12); 4.13 (2H, q,J=7.13); 2.30 (4H, m); 1.93 (1H, m); 1.27 (3H, t, J=7.1), 0.95 (3H, s);0.80 (3H, s); 0.68 (3H, s) ppm.

EXAMPLE 49

Synthesis of 6.27

Aldehyde 6.26 is coupled with 13.1 as described for analogue 11 from6.12 (yield 91% with Rf 0.73, Et₂O:hexane 1:1).

¹H NMR: (360 MHz, CDCl₃): δ: 6.34 (1H, dd, J=11, 15); 5.92 (1H, d,J=11); 5.66 (dt, J=8,15); 5.20 (1H, br s); 4.87 (1H, br s); 4.39 (1H, t,J=5.5); 4.185 (1H, m); 4.13 (2H, q, J=7.14); 2.40 (1H, dd, J=3, 13);2.30 (2H, m); 2.18 (1H, dd, J=7, 13); 1.26 (3H, t, J=7.14); 0.882 (9H,s); 0.866 (9H, s); 0.80 (3H, s); 0.78 (3H, s); 0.66 (3H, s); 0.07 (12H,s) ppm.

EXAMPLE 50

Synthesis of 6.29

To a suspension of copper(I)iodide (420 mg, 2.2 mmol) and zinc dust (600mg, 9.2 mmol) in ethanol-water 7:3 (27 ml) are addedtrans-2,4-pentadianoic acid ethyl ester (270 μl, 1.93 mmol), and iodide6.2 (420 mg, 1.5 mmol). The mixture is sonicated during 1 h under argonat 0° C. The mixture is filtered through celite and washed with EtOAc.The filtrate is extracted with EtOAc, dried (MgSO₄) and concentrated.Column chromatography (silica gel:diethyl ether:hexane 1:9→1:5) gives6.28 (145 mg, 35%) and recovered 6.2 (145 mg, 35%).

Rf: 0.38 (diethyl ether:hexane 1:1).

¹H NMR: (500 MHz, CDCl₃): δ: 5.55 (2H, 2×dt, J=6, 15); 4.14 (2H, q,J=7.13); 3.58 (1H, br d, J=11); 3.45 (1H, br d, J=11); 3.02 (2H, d,J=6); 2.10 (1H, m); 1.90 (2H, m); 1.75 (1H, qd, J=3,10); 1.26 (3H, t,J=7.12); 0.98 (3H, s); 0.89 (3H, s); 0.72 (3H, s) ppm.

To a solution of 6.28 (40 mg, 142 μmol) in dry EtOAc (8 ml), a catalyticamount of Pd/C (10%) is added, after which the mixture is hydrogenatedfor 3 h (4 atm). Filtration over celite, addition of Et₃N (200 μl),evaporation and column chromatography (diethyl ether:hexane 1:4) gives6.29 [32 mg, 80%, with Rf=0.36 (diethyl ether:hexane 1:1)].

¹H NMR: (500 MHz, CDCl₃): δ: 4.13 (2H, q, J=7); 3.58 (1H, d, J=10); 3.46(1H, d, J=10); 2.30 (2H, t, J=7); 1.87 (1H, m); 1.72 (1H, qd, J=3, 10);1.26 (3H, t, J=7); 0.99 (3H, s); 0.89 (3H, s); 0.71 (3H, s) ppm.

EXAMPLE 51

Synthesis of 10.2

A solution of 10.1 (3.44 g, 17.36 mmol), ethylene glycol (5.3 ml, 95mmol) and pyridinium p-toluenesulfonate (500 mg, 1.99 mmol) incyclohexane (190 ml) is refluxed for 3 h with continuous separation ofwater. After cooling to r.t., the solvent is evaporated, and the residueis dissolved in diethyl ether (300 ml). Washing with a saturatedNaHCO₃-solution and brine, drying (Na₂SO₄), solvent evaporation andpurification by column chromatography (silica gel hexanes:acetone 9:1)and HPLC (isooctane:acetone 95:5) gives 10.2 (3.6 g, 86%).

Rf: 0.20 (hexane:acetone 95:5).

IR (film): 2950; 2881; 1740; 1436; 1280; 1189 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 3.93 (4H, m); 3.65 (3H, s); 2.47 (1H, dd,J=14.45, 3.13); 2.06 (1H, tt, J=11.15, 3.31); 2.02 (1H, dd, J=14.42,10.72); 1.63-1.52 (5H, m); 1.22 (1H, m); 0.911 (s), 0.898 (3H, s) ppm.

EXAMPLE 52

Synthesis of 10.3

To a stired solution of LDA (2M in hexane, 4.67 ml, 9.348 mmol) in THF(5.45 ml) at −30° C. is added a solution of 10.2 (1.510 g, 6.232 mmol)in THF (21.8 ml), and stirring is continued for 1 h. After cooling to−78° C., a mixture of 5-bromo-1-pentene (2.34 ml, 19.76 mmol) andhexamethylphosphoramide (5.5 ml, 31.16 mmol) is added; stirring iscontinued for 3 h. The mixture is allowed to come very slowly to r.t.and is then diluted with water and diethyl ether. Extraction of thewater layer with diethyl ether, drying of the organic phase (Na₂SO₄,solvent evaporation and purification by column chromatography (silicagel:hexane:acetone 9:1) and HPLC (isooctane:acetone 97:3) yields 10.3(1.81 g, 93%).

Rf: 0.24 (isooctane:acetone 97:3).

IR (film): 3076; 2950; 2881; 1732; 1641; 1435; 1186 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 5.76 (1H, ddt, J=17.10, 10.17, 6.65 (t));4.99 (1H, ddd, J=17.13, 1.82, 1.59); 4.94 (1H, m); 3.92 (4H, m); 3.65(3H, s); 2.49 (1H, dt, J=11.61, 3.39 (t)); 2.03 (2H, m); 1.95 (1H, dt,J=12.87, 3.52(t)); 1.65-1.59 (3H, m); 1.57 (1H, m); 1.53 (1H, m);1.51-1.40 (3H, m); 1.35-1.19 (3H, m); 0.961 (3H, s); 0.924 (3H, s) ppm.

EXAMPLE 53

Synthesis of 10.6

To a suspension of LiAlH₄ (332.5 mg, 8.762 mmol) in diethyl ether (165ml) is added dropwise at 0° C. a solution of 10.3 (1.600 g, 5.154 mmol)in diethyl ether (82 ml); the mixture is stirred for 1 h at 0° C. andfor 3 h at r.t. To the vigorously stirred mixture is then added, veryslowly, a saturated Na₂SO₄-solution, until a white precipitateflocculates. The suspension is stirred for 1 h, the precipitate filteredover celite, and the solvent evaporated, yielding 10.4 (1.453 g, 99.8%).

Rf: 0.22 (hexane:acetone 8:2).

¹H NMR: (500 MHz, CDCl₃): δ: 5.79 (1H, ddt, J=17.10, 10.17, 6.64(t));4.99 (1H, ddd, J=17.14, 1.94, 1.61); 4.94 (1H, m); 3.92 (4H, m); 3.55(1H, m); 3.40 (1H, m); 2.03 (2H, m); 1.77 (1H, dd, J=12.82, 3.55); 1.63(1H, m); 1.57 (1H, dd, J=12.72, 3.67); 1.54-1.28 (12H, m); 0.978 (3H,s); 0.910 (3H, s) ppm.

To a solution of 10.4 (1.453 g, 5.145 mmol) in dichloromethane (25.7 ml)and triethylamine (3.9 ml, 20.58 mmol), a solution of TsCl (1.962 g,10.29 mmol) in dichloromethane (15.4 ml), and a small amount of4-dimethylaminopyridine are added at 0° C. After stirring for 20 h atr.t. the volume is reduced to 50%, followed by filtration of theprecipitate. Complete evaporation of the solvent, and HPLC purification(hexane:acetone 85:15) gives 10.5 (2.126 g, 95%).

Rf: 0.25 (hexane:acetone 85:15).

To a solution of 10.5 (2.126 g, 4.870 mmol) in diethyl ether (250 ml) isadded LiAlH₄ (3.69 g, 97.40 mmol) and the refluxed suspension is stirredfor 5 h. After cooling to 0° C. a saturated Na₂SO₄-solution is carefullyadded until the grey precipitate has disappeared. A small excess ofNa₂SO₄-solution is added and stirring is continued for 3 h. Theprecipitate is filtered over celite and is washed twice by suspending itin diethyl ether, followed by a new filtration. After evaporating thesolvent, the residue is purified by HPLC (isooctane:ethyl acetate 98:2),giving 10.6 (1.14 g, 88%).

Rf: 0.20 (isooctane:ethyl acetate 98:2).

IR (film): 3076; 2949; 2869; 1641; 1464; 1090 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 5.81 (1H, ddt, J=17.17, 10.21, 6.63(t));4.99 (1H, ddd, J=17.02, 2.08, 1.58); 4.93 (1H, m); 3.92 (4H, m); 2.02(2H, m); 1.64 (2H, m); 1.54 (2H, m); 1.52-1.33 (7H, m); 1.26 (2H, m);0.971 (3H, s); 0.911 (3H, s); 0.907 (3H, d, J=6.83); 0.895 (1H, m) ppm.

EXAMPLE 54

Synthesis of 10.7

At −78° C., ozone is passed through a solution of 10.5 (565 mg, 2.121mmol) in dichloromethane (16.8 ml) and a 2.5M solution of sodiumhydroxide in methanol (4.24 ml), until a light blue color is retained.The reaction mixture is diluted with diethyl ether and water. After thetemperature has raised to room temperature, the organic phase is washedwith brine, dried (Na₂SO₄) and the solvent is evaporated. Purificationof the residue by column chromatography (hexane:acetone 9:1) and HPLC(hexane:acetone 97:3) gives the ester (405 mg, 64%). A solution of thisester (400 mg, 1.340 mmol) and pyridinium p-toluenesulfonate (101 mg,mmol) in acetone 13.4 ml), and a few drops of water, is refluxed for 3h. After cooling to r.t., the solvent is evaporated and the residue isdissolved in diethyl ether, followed by washing with a saturatedNaHCO₃-solution and brine. Drying (Na₂SO₄), solvent evaporation andpurification by column chromatography (silica gel:hexane:acetone 9:1),and HPLC (hexane:acetone 96:4), gives 10.7 (256 mg, 75%) next to 10.6(83 mg, 21%).

Rf: 0.19 (hexane:acetone 93:7).

IR (film): 1739; 1705 (s); 1454; 1436; 1249 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 3.67 (3H, s); 2.49 (1H, dt, J=13.62(t),6.39); 2.33-2.24 (3H, m); 2.03 (1H, ddd, J=12.90; 6.24; 3.29); 1.80-1.58(4H, m); 1.56-1.47 (3H, m); 1.33 (1H, m); 1.11 (3H, s); 1.07 (3H, s);1.02 (1H, m); 0.943 (3H, d, J 6.90) ppm.

EXAMPLE 55

Synthesis of Aldehydes 10.8 and 10.9

A. To a solution of FOSMIC (19.6 μl, 113.6 μmol) in diethyl ether (475μl) is added at −60° C. a 2.5M solution of butyllithium in hexanes (52μl, 130.5 μmol), and the resulting solution is stirred for 15 minutes. Asolution of 10.7 (28.9 mg, 113.6 μmol) in diethyl ether (119 μl) is thenadded, and the mixture is allowed to come to 0° C. and stirring iscontinued for 1.5 h. After adding, carefully, a 37%-aquous HCl solution(200 μl) the mixture is vigorously stirred overnight. After dilutingwith diethyl ether, the water layer is extracted with diethyl ether andethyl acetate, followed by washing the organic phase with brine anddrying (Na₂SO₄). The solution is treated with diazomethane, the excessis destroyed by adding silica gel. Filtration, solvent evaporating andpurification by column chromatography (silica gel:hexane:acetone 9:1)gives 10.8 and 10.9 (7.4:1 ratio; 17 mg, 64%). The mixture can beseparated by HPLC (hexane:acetone 96:4).

Rf: 0.35 (hexane:acetone 9:1).

IR (film): 2934; 2863; 1739; 1719; 1438; 1374; 1248; 1171 cm⁻¹.

hu 1H NMR: (500 MHz, CDCl₃): δ: 9.80 (1H, d, J=2.89); 3.66 (3H, s); 2.29(2H, m); 1.99 (1H, dt, J=12.53, 3.29(t)); 1.86 (1H, m); 1.74 (2H, m);1.63-1.44 (4H, m); 1.39 (1H, m); 1.23 (2H, m); 1.17 (3H, s); 1.02 (1H,m) ppm.

B. A suspension of trimethylsulfonium iodide (107.0 mg, 0.514 mmol) and2.5M solution of butyllithium (in hexane 132 μl, 0.29 mmol) in THF (6.2ml) is stirred for 1 h at r.t. After cooling to 0° C., a solution of10.7 (52.3 mg, 0.206 mmol) in THF (4.1 ml) is added and stirring iscontinued for 2 h at. r.t. The mixture is diluted with dichloromethane,extracted with water and brine, dried (Na₂SO₄), and the solventevaporated. The residue is purified by column chromatography (silicagel:hexane:acetone 9:1) and HPLC (hexane:acetone 96:4), yielding the twodiastereoisomers 10.10 (17 mg, 33% ratio 6:4) and starting material 10.7(18 mg, 33%).

Rf: 0.17 (hexane:acetone 96.5:3.5).

To a solution of 10.7 (17 mg, 63.34 μmol) in diethyl ether (3.2 ml) isadded at 0° C. boron trifluoride diethyl etherate (40 μl, 324.6 μmol).The solution is stirred for 1 h at 0° C. and 12 h at r.t.; The mixtureis poured in diethyl ether and washed with a saturated NaHCO₃-solutionand brine. After evaporation of the solvent the residue is purified bycolumn chromatography (silica gel:hexane:acetone 9:1), yielding amixture of 10.8 and 10.9 (11 mg, 65%; ratio 1.6:1). The mixture can beseparated by HPLC (hexane:acetone 96:4).

Rf: 0.38 (hexane:acetone 9:1).

IR (film): 2950; 2867; 1739; 1713; 1437 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 9.98 (1H, d, J=2.60); 3.67 (3H, s); 2.30(2H, m); 2.01 (1H, m); 1.84 (1H, m); 1.77-1.64 (4H, m); 1.55-1.28 (6H,m); 1.18 (3H, s); 1.01 (3H, s); 0.941 (1H, m); 0.927 (3H, d, J=6.88)ppm.

EXAMPLE 56

Synthesis of 11.13

A solution of (−)-camphoric acid 11.12 (3 g, 15 mmol) in dry THF (45 ml)is added slowly to a stirred suspension of LiAlH₄ (1.9 g, 50 mmol) indry Et₂O (40 ml); the mixture is refluxed for 4 h. After cooling tor.t., Na₂SO₄.10 H₂O is added. Filtering, solvent evaporation andcrystallisation from EtOAc yields the diol (2.28 g, 88%).

A solution of the diol (0.54 g, 3.14 mmol) in vinyl acetate (10 ml) istreated for 66 h with SAM II lipase (300 mg) at 37° C.

Solvent evaporation and column chromatography (silicagel, pentane:EtOAc8:2) yields monoacetate 11.13 (0;4 g, 60%).

Rf: 0.28 (pentane:EtOAc 8:2).

IR (film): 3440 (s, broad); 2962 (s); 2874 (m); 1739 (s); 1463 (m); 1369(m); 1246 (s); 1144 (w); 1033 (s); 971 (w) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 4.08 (1H, dd, J=10.80); 3.98 (1H, dd,J=6.10, 8.20); 3.58 (1H, d, J=10.75); 3.46 (1H, d); 2.20 (1H, ddt,J=8.9); 2.03 (3H, s); 1.89 (1H, ddd), 1.58 (1H, dt); 1.35 (2H, 2); 1.01(6H, s); 0.81 (3H, s) ppm.

EXAMPLE 57

Synthesis of 11.14

To 11.13 (317 mg, 1.48 mmol) and Et₃N (1.71 ml, 14.8 mmol) inCH₂Cl₂:DMSO (1:1; 8 ml) is added SO₃.pyridine complex (1.42 g, 8.88mmol) After stirring for 3 h at r.t. the mixture is poured in H₂O andextracted with Et₂O. The organic layer is washed with 1 N HCl and withbrine, dried (Na₂SO₄) and concentrated. Column chromatography(silicagel, pentane:EtOAc 9:1) gives the aldehyde (250 mg, 80%).

This aldehyde (190 mg, 0.90 mmol) in dry THF (2 ml) is added to lithiotriethyl-4-phosphonoacetate (2.83 mmol; from phosphonoacetate and LDA)in dry THF (8 ml) at 0° C. After stirring for 12 h at 25° C., themixture is washed with brine and dried (MgSO₄). Solvent evaporationyields crude acetate which is solvolysed with K₂CO₃ in EtOH at r.t.Filtration, solvent evaporation and column chromatography (silicagel;pentane:EtOAc 75:25) yields 11.14 (155 mg, 65%).

Rf: 0.31 (pentane:EtOAc 8:2).

IR (film): 3436 (s, broad); 2965 (s); 2870 (m); 1712 (s); 1636 (s); 1462(s); 1369 (s); 1330 (m); 1253 (s); 1140 (s); 1007 (s); 882 (m); 832 (w)cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 7.28 (1H, dd, J 10.5, 15.4); 6.18 (1H, d,J=15.5); 6.10 (1H, dd); 5.80 (1H, d); 4.19 (2H, q, J=7.2); 3.71 (1H, dd,J=10.3, 5.8); 3.53 (1H, dd, J=8.2); 2.13 (1H, m); 2.00 (1H, m); 1.94(1H, m); 1.48 (1H, m); 1.41 (1H, m); 1.28 (3H, t); 1.01 (3H, s); 0.94(3H, s); 0.68 (3H, s) ppm.

EXAMPLE 58

Synthesis of 11.16

A solution of 11.14 (17 mg, 0.064 mmol) in EtOAc (1 ml), and 5% Rh onAl₂O₃ (20 mg) is stirred at r.t. for 2 h under H₂ atmosphere. Filtrationon silicagel, solvent evaporation and HPLC (pentane:EtOAc 7:3)purification gives 11.16 (16 mg, 90%).

Rf: 0.29 (pentane:EtOAc 75:25).

IR (film): 3385 (s, broad); 2941 (s); 2860 (m); 1735 (s); 1455 (s); 1371(s); 1248 (s); 1152 (s); 1022 (s); 945 (m); 870 (w) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.12 (2H, q, J=7.1); 3.72 (1H, dd, J=10.2,5.4); 3.50 (1H, dd, J=8.7); 2.30 (2H, t, J=7.4); 2.07 (1H), 1.88 (1H);1.60 (2H, m); 1.53 (1H); 1.40 (1H); 1.38-1.18 (5H, m); 1.25 (3H, t);0.89 (3H, s); 0.84 (3H, s); 0.68 (3H, s) ppm.

EXAMPLE 59

Synthesis of 11.18

A solution of 11.16 (10 mg, 0.037 mmol) in dry Et₂O (1 ml) and EtMgCl(2M sol. in Et₂O, 144 μl, 289 μmol) is stirred for 90 min at r.t. Onedrop of saturated NH₄Cl solution is then added. Filtration on silicagel,rincing with pentane:EtOAc (6:4), solvent concentration and HPLC(pentane:EtOAc 6:4) purification gives 11.18 (7.8 mg, 75%).

Rf: 0.20 (pentane:EtOAc 8:2).

IR (CH₂Cl₂): 3349 (s, broad); 2966 (s); 2936 (m); 2874 (m); 1457 (m);1388 (m); 1374 (m); 1264 (w); 1094 (m); 1034 (m); 973 (w); 946 (w); 878(w) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 3.72 (1H, dd, J=5.41, 0.1); 3.51 (1H, dd,J=8.7); 2.09 (1H, dddd, J=9.6); 1.90 (1H, dddd, J=13); 1.46 (4H, q,J=7.4); 1.65-1.15 (11H, m); 0.90 (3H, s); 0.86 (6H, t); 0.84 (3H, s);0.69 (3H, s) ppm.

EXAMPLE 60

Synthesis of 11.17

From 11.16 and MeMgBr as described for 11.18 (yield 86%)

Rf: 0.37 (pentane:EtOAc 5:5).

IR (CH₂Cl₂): 3354 (s, broad); 2937 (s); 2868 (m); 1466 (s); 1375 (s);1204 (w); 1150 (w); 1090 (w); 1040 (m); 1008 (m); 905 (w) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 3.72 (1H, dd, J=5.15, 9.85); 3.51 (1H, dd,J=9.3); 2.08 (1H, m); 1.89 (1H, m); 1.53 (1H, m), 1.50-1.15 (10H, m);1.21 (6H, s); 0.90 (3H, s); 0.85 (3H, s); 0.69 (3H, s) ppm.

EXAMPLE 61

Synthesis of 11.19

To a solution of 11.17 (12 mg, 47 μmol), N-methyl morfoline oxide (8.5 mg, 72 μmol) and activated molecular sieves (4 Å; 24 mg) in CH₂Cl₂ (400μl) is added tetra-n-propyl ammonium perruthenate (0.8 mg, 2.35 μmol).After stirring for 2 h at r.t. the mixture is filtered on silicagel. Theresidue is washed with pentane:EtOAc 5:5. Solvent evaporation and HPLC(pentane:EtOAc 85:15) purification gives 11.19 (8;3 mg, 70%).

Rf: 0.38 (pentane:EtOAc 8:2).

IR (CH₂Cl₂): 3421 (s, broad); 2966 (s); 2862 (m); 2718 (w); 1713 (s);1466 (s); 1377 (s); 1133 (w); 905 (w) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 9.76 (1H, d, J=2.3); 2.70 (1H); 2.04 (1H),1.75-1.21 (1H, m); 1.21 (6H, s); 1.09 (3H, s); 0.87 (3H, s); 0.84 (3H,s) ppm.

EXAMPLE 62

Synthesis of 11.20

From 11.18 as described for 11.19 from 11.17 (yield 78%).

Rf: 0.15 (hexane:ethyl acetate 85:15).

IR (film): 3442 (s, broad); 2938 (s); 2871 (m); 2720 (w); 1717 (s); 1457(s); 1377 (s); 1262 (m); 1092 (m); 1031 (s); 947 (w); 877 (w) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 9.76 (1H, d, J=2.2); 2.70 (1H, ddd); 2.06(1H, m); 1.74-1.59 (2H, m); 1.51 (1H, m); 1.46 (4H, q, J=7.5); 1.41 (1H,m); 1.39-1.20 (1H, m); 1.09 (3H, s); 0.87 (3H, s); 0.86 (6H, t); 0.84(3H, s) ppm.

EXAMPLE 63

Synthesis of 11.23

To a stirred solution of LDA (2.11 mmol) THF (2 ml) is added at −78° C.,a solution of 11.22 (0.58 g, 1.85 mmol) in THF (0.4 ml) over a period of10 minutes. The resulting solution is warmed up slowly to r.t. Afterstirring at r.t. for 2 h the reaction mixture is cooled to −78° C. andPhNTf₂ (0.71 g, 2.0 mmol) in THF (2.5 ml) is added dropwise. Thesolution is warmed up slowly to 0° C. and stirred overnight. Water isadded extraction with pentane, drying (MgO₄) and solvent evaporation andpurification by column chromatography (silicagel, hexane:EtOAc 10:1)gives 11.23 (0.47 g, 65%).

Rf: 0.30 (hexane:EtOAc 10:1).

IR (film): 3750; 2973; 1417; 1209; 114 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 5.60 (1H, dd, J=6.86, 3.50); 4.90 (1H, q,J=5.3); 3.54 (1H, m); 3.48 (1H, m); 2.5 (1H, m); 2.3 (2H, m); 2.0 (2H,m); 1.8 (1H, m); 1.5-1.4 (10H, m); 1.33 (3H, d, J=5.28); 1.21 (3H, s);1.19 (3H, s); 1.18 (3H, t, J=7.05); 1.05 (1H, m); 0.94 (3H, d, J=6;52);0.76 (3H, s) ppm.

EXAMPLE 64

Synthesis of 11.24

Through a solution of 11.23 (400 mg, 0.83 mmol) and solid NaHCO₃ (112mg, 1.3 mmol) in MeOH (200 ml) is passed a stream of ozone (18 mmol/h),generated by a WELSBACH generator, at −78° C. over a period of 30 minwhile the solution turned to deep blue. The solution is then flushedwith nitrogen until the solution became colourless. NaBH₄ (1.0 g, 26mmol) is added to the mixture at −78° C., after 15 min, another portion(1.0 g, 26 mmol) is added. The mixture is allowed to warm up slowly tor.t. and is stirred for 18 h. Sodium borohydride (2.0 g, 52 mmol) isadded at −20° C. and the reaction mixture is stirred for 2 h and thenslowly warmed up to r.t. MeOH is evaporated and saturated NH₄Cl solutionis added. Extraction with CH₂Cl₂, drying (MgSO₄) and solvent evaporationgives 11.24 (300 mg, 91%).

Rf: 0.23 (hexane:EtOAc 2:1).

IR (film): 3397, 2970; 2348; 1713; 1416; 1209; 114; 904 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.90 (1H, q, J=5.3); 3.65 (3H, s); 3.64(2H, m); 3.54 (1H, m); 3.48 (1H, m); 2.70 (1H, t, J=9.36); 2.0 (1H, m);1.80 (1H, m); 1.70 (2H, m); 1.5-1.4 (7H, m); 1.30 (6H, m); 1.20 (10H,m); 1.10 (1H, m); 1.0 (3H, t, J=6.43); 0.80 (3H, s) ppm.

EXAMPLE 65

Synthesis of 11.26

To a solution of 11.24 (90 mg, 0.2 mmol) is added a solution of TsCl(167 mg, 0.87 mmol) in pyridine (2.5 ml). The mixture is stirred at −4°C. for 18 hrs. An ammonium acetate solution is added, extraction withCH₂Cl₂, drying (MgSO₄) and solvent evaporation gives a crude oil whichis used in the next reaction. To LiAlH₄ (160 mg, 4.2 mmol) in dry THF (5ml), is added 11.25 (460 mg, 0.11 mmol) in dry THF (5 ml) at 0° C. Themixture is refluxed for 36 h, then 10% HCl solution is carefully addeduntil neutralization. Solvent evaporation and HPLC purification(hexane:EtOAc 1:1) gives 11.26 (31 mg, 61%).

Rf: 0.29 (hexane:EtOAc 1:1).

IR (film): 3855; 2957 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 3.72 (1H, dd, J=10.20, 4.70); 3.41 (1H, dd,J=10.20, 9.07); 1.92 (1H, m); 1.70 (2H, m); 1.60 (1H, s br); 1.56 (1H,m); 1.5-1.3 (12H, m); 1.25 (1H, m); 1.20 (6H, s); 1.04 (1H, m); 0.95(3H, d, J=6.71); 0.85 (3H, t, J=7.15); 0.67 (3H, s) ppm.

EXAMPLE 66

Synthesis of 11.27

To a mixture of 11.26 (30 mg, 100 μmol), N-methylmorfolin oxide (1.66eq, 166 μmol, 19 mg) and molecular sieves (54 mg, type 4 Å, 2 à 3μ) inCH₂Cl₂ (1.5 ml), tetrapropylammonium perruthenate (5 μmol, 1.8 mg) isadded. After stirring for 1 h the greyblack suspension is purified bydirect column chromatography (Et₂O:hexane 1:4→1:1) yielding 11.27 (15mg, 50%).

Rf: 0.33 (Et₂O:hexane 1:1).

¹H NMR: (500 MHz, CDCl₃): δ: 9.69 (1H, d, J=3.31); 2.58 (td, J=3.3,9.1); 1.98 (1H, m); 1.87 (1H, m); 1.22 (6H, s); 0.96 (3H, d, J=6.68);0.92 (3H, t, J=7.2); 0.87 (3H, s) ppm.

EXAMPLE 67

Synthesis of 12.2

A mixture of 12.1 (75 mg, 0.268 mmol) and sodium methoxide (catalyticamount) in super dry methanol (1.5 ml) is stirred for 24 hrs at r.t.under Ar. The mixture is then filtered through silicagel, the filtrateconcentrated in vacuo and separated by HPLC (silicagel; hexane:ethylacetate 75:25) to yield the cis fused ketone (55 mg, 73%) next to thetrans isomer (18 mg).

The cis ketone (50 mg, 0.179 mmol) and 1-(trimethylsilyl)imidazole (104μl, 0.716 mmol) in dichloromethane (1.8 ml) is stirred for 3 hrs at roomtemperature. After solvent removal in vacuo, diethylether is added andthe resulting precipitate is filtered off on a short silica pad.Concentration of the filtrate yields 73 mg of a crude product which ispurified on HPLC silicagel;hexane:ethyl acetate 95:5) to give theprotected alcohol 12.2 (50 mg, 79%).

Rf: 0.58 (hexane:ethyl acetate 85:15).

IR (film): cm⁻¹.2958 (s), 1710 (s), 1464 (m), 1379 (m), 1320 (w), 1249(s), 1156 (m) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 2.31 (3H, m), 2.15 (1H, m), 1.95-1.70 (5H,m), 1.57 (1H, m), 1.43-1.25 (8H, m), 1.18 (6(+1)H, s), 1.04 (3H, s),0.91 (3H, d), 0.10 (9H, s) ppm.

EXAMPLE 68

Synthesis of 12.5

A suspension of NaH (956 mg; 23.9 mmol) in anhydrous dimethyl sulfoxide(30 ml) is stirred at 65° C., under nitrogen, for 1.5 h, after which3-ethoxyethyl-3-methyl-1-butyn (3.68 g; 23.9 mmol) is slowly added. Asolution of the 12.3 (1.77 g; 4.84 mmol) in dry dimethylsulfoxide (10ml) is then added and stirring is continued for 0.5 h at r.t. Thereaction mixture is poured into an ice cold saturated NH₄Cl solution.The aquous phase is extracted with ether, and the combined extracts arewashed with brine, dried (MgSO₄) and evaporated under reduced pressure.The residue is purified by column chromatography (silicagel;hexane:ethyl acetate 8:2) to give 12.4 (1.24 g; 67% yield) of theproduct. A mixture of this alcohol (560 mg; 1.6 mmol) and pyridiniumdichromate (1.8 g; 4.8 mmol) in dichloromethane (10 ml) is stirred for 2hrs at r.t. The resulting ketone is directly purified by columnchromatography (hexane: ethyl acetate 8:2); 467 mg (84%) is obtained.

A solution of this ketone (369 mg; 1.06 mmol) and a catalytic amount ofsodium methoxide in dry methanol (10 ml) is stirred under nitrogen atr.t. for 12 hrs. The reaction mixture is filtered on silicagel, usingmethanol as the eluent. Evaporation under reduced pressure gave aresidue that was purified on a silicagel column (ethyl acetate:hexane2:8). Pure 12.5 (149 mg; 65%) is obtained upon separation by HPLC (ethylacetate:hexane 2:8).

Rf: 0.48 (hexane:ethyl acetate 8:2).

IR (film): 3398, 2979, 2934, 2875, 2291, 2226, 1708, 1464, 1443, 1378,1360, 1334, 1253, 1160, 1124, 1081, 1053 cm⁻¹.

¹H NMR: (200 MHz, CDCl₃): δ: 1.18 (3H, t); 3.67 (lH, dq, J=9.11 7.05Hz); 3.49 (1H, dq, J=9.11, 7.10 Hz); 5.68 (1H, q, 5.24 Hz); 1.32 (3H, d,5.24 Hz); 1.49 (3H, s); 1.43 (3H, s); 2.23 (2H, m); 1.06 (3H, d, 6.48Hz); 1.06 (3H, s); 2.44 (1H, m) ppm.

EXAMPLE 69

Synthesis of 12.6

A solution of 12.4 (1.035 g; 3.1 mmol) and p-toluene sulfonic acid (295mg; 1.55 mmol) in toluene (50 ml) is stirred at 60° C. for 1 h. Thereaction mixture is then poured in saturated NaHCO₃ solution, extractedwith diethylether, washed and dried (MgSO₄). Column chromatography(silicagel, hexane-ethyl acetate 85:15) of the residue, obtained uponfiltration and solvent evaporation gives 12.6 (567 mg, 70%).

Rf: 0.45 (hexane:ethyl acetate 8:2).

IR (film): 3426, 2932, 2868, 2223, 1615, 1457, 1372, 1165 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 5.14 (1H, s, J=28.78 Hz); 5.20 (1H, s);1.88 (3H, s); 2.34 (1H, d, J=3.41 Hz); 2.38 (1H, d, J_(c′d)=3.45 Hz,J_(cc′)=16.76 Hz); 1.06 (3H, d, J=6.56 Hz); 0.93 (3H, s); 4.08 (1H, m)ppm.

EXAMPLE 70

Synthesis of 12.7

A mixture of 12.6 (300 mg; 1.15 mmol), 3-chloro-peroxybenzoic acid (497mg, 80%; 2.31 mmol and Na₂HPO₄ (163 mg; 1.15 mmol) in dry THF (30 ml) isstirred at 0° C. under nitrogen atmosphere. After 3 days stirring, themixture is diluted with ethyl acetate:hexane (1:1), washed with 10%Na₂SO₃ solution, with saturated Na₂CO₃ solution and with brine and isdried (MgSO₄). Filtration and removal of the solvents under reducedpressure, and column chromatography (silicagel;ethyl acetate:hexane 2:8)gives the epoxide (90 mg; 66%).

A mixture of this product (50 mg; 0.181 mmol) and pyridinium dichromate(203 mg; 0.54 mmol) in dichloromethane (4 ml) is stirred for 2 h at r.t.The reaction mixture is purified by column chromatography (hexane:ethylacetate 8:2) to give the trans fused ketone (36 mg; 69%).

A solution of the ketone (80 mg; 0.328 mmol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (76 mg; 0.500 mmol) in dichloromethane (2 ml) isstirred at r.t. for 3 days. The reaction mixture is poured in saturatedNH₄Cl solution, extracted with diethylether washed with saturated NaHCO₃and brine, and dried (MgSO₄). The residue, after filtration and removalof the solvents, is purified on HPLC (ethyl acetate:hexane 15:85) andaffords 12.7 (8 mg, 27%) next to the trans fused isomer (30 mg).

Rf: 0.24 (ethyl acetate:hexane 15:85).

IR (film): 3410, 2952, 2239, 1712, 1460, 1379, 1139, 1307, 1271, 1232,1152, 1068 cm⁻¹.

¹H NMR: (200 MHz, CDCl₃): δ: 1.52 (3H, s); 2.95 (1H, d, 4.47 Hz); 2.71(1H, d, 5.65 Hz); 1.05 (3H, s); 1.05 (3H, d, 6.5 Hz) ppm.

EXAMPLE 71

Synthesis of 12.10

Ozonolysis of vitamin D₂ (2 g; 5.05 mmol) in dichloromethane:methanol 50ml, (1:1) is carried out at −78° C. Subsequent work-up withdimethylsulfide (8 ml) at −78° C. for 30 min and evaporation of thesolvent gives crude 12.8. It is dissolved in tetrahydrofuran (30 ml) and5% HCl (10 ml) is added under stirring. Stirring is continued at 30° C.,under nitrogen, for 36 hrs. Evaporation of the solvent, addition ofdiethylether, washing with saturatted NaHCO₃ solution, drying (MgSO₄)and concentration in vacuo affords a residue. Flash chromatography(silicagel; hexane:ethyl acetate 8:2) gives white crystalline 12.9together with the 20-S-isomer 736 mg, 70%; 2.5:1 ratio). This mixture(200 mg; 0.96 mmol) in methanol (25 ml) is treated with NaBH₄ (73 mg,1.92 mmol) at r.t., under N₂, for 20 min. A HCl solution (10%, 8 ml) isadded, after stirring for 10 min, the methanol is evaporated. Additionof diethylether, washing with saturated NaHCO₃, drying (MgSO₄)evaporation and separation of the C-20 epimers (HPLC; hexane:ethylacetate:methanol; 100:100; 1.5) yields 12.10 (140 mg; 69%).

Rf: 0.45 (hexane:ethyl acetate:methanol 5:4:1).

IR (film): 2795, 2703, 1719, 1700, 1380 cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 0.86 (3H, d, J=6.75 Hz) 0.89 (s) and 1.02(s) (3H), 3.30-3.94 (3H, m) ppm.

EXAMPLE 72

Synthesis of 12.12

A solution of 12.10 (170 mg, 0.8 mmol) and TsCl (229 mg, 1.20 mmol) indry pyridine (10 ml) is kept at 0° C. for 13 hrs. The mixture is thenpoured in ice-water, extraction, washing (NaHCO₃ sat. solution) drying(MgSO₄) solvent evaproation and flash chromatography (silicagel;hexane:ethyl acetate 6:4) gives 12.11 (163 mg, 56%). Reaction of 12.11(160 mg, 0.44 mmol) with the anion of 3-ethoxyethyl-1-butyn (2.2 ml) asdescribed for 12.4, in example 61, gives after work-up and flashchromatography (silicagel;hexane:ethyl acetate 8:2) 86 mg (56%) of theproduct. A mixture of this alcohol (56 mg, 0.16 mmol) and pyridiniumdichromate (241 mg, 0.64 mmol) in dry dichloromethane (7 ml) is stirredat r.t., under N₂ for 1 h. Direct flash chromatography (silicagel;hexane:ethyl acetate 8:2) gives 12.12 (39 mg; 70%).

Rf: 0.21 (hexane:ethyl acetate 9:1).

IR (film): 2231, 1708 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 0.95 (d, J=6.56 Hz) and 0.96 (d, J=6.67 Hz)(3H), 1.03 (3H, s), 1.18 (3H, m), 1.31 (3H, d, J=5.31 Hz), 1.42 (3H, s),1.478 (s) and 1.482 (s) (3H), 3.43-3.69 (2H, m), 5.08 (1H, m) ppm.

EXAMPLE 73

Synthesis of 12.13

From 12.10 and 3-(ethoxy)-ethoxy-ethyl-1-pentyne as described for 12.12.

Rf: 0.35 (hexane:EtOAc ˜9:1).

IR (film): 2234; 1708 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 5.10 (1H, q, J 5.21); 3.68 (1H, m); 3.48(1H, m); 1.31 (3H, d, J=5.21); 1.17 (3H, dd, J=7.03, 7.03); 1.04 (3H,s); 0.97 (3H, d, J=6.63); 0.94 (6H, m) ppm.

EXAMPLE 74

Synthesis of 12.14

A mixture of 12.12 (19 gm, 0.055 mmol), 5% Rh/Al₂O₃ (8 mg) and EtOAc(2.5 ml) is stirred at r.t. uner H₂ (atmospheric pressure) for 1 h. Themixture is filtered through a short silica gel column (hexane:EtOAc7:3). HPLC purification (hexane:EtOAc 9:1) gives 12.14 (17 mg, 89%).

Rf: 0.50 (hexane:EtOAc 8:2).

IR (film): 1708 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.86 (1H, q, J=5.33); 3.50 (2H, m); 1.26(3H, d, J=5.33); 1.19 (3H, s); 1.17 (3H, dd, J=6.90, 6.90); 1.17 (3H,s); 1.02 (3H, s); 0.82 (3H, d, J=6.67) ppm.

EXAMPLE 75

Synthesis of 12.15

To a solution of LDA (0.50 mmol) in dry THF (3.5 ml) at −78° C., underN₂, triethyl 4-phosphonocrotonate (90%, 124 μl, 0.50 mmol) is addeddropwise. Stirring is continued at −78° C. for 30 min. A solution of12.9 (88 mg, 0.42 mmol) in dry THF (1.5 ml) is added dropwise. Thereaction is stirred at −78° C. for 2 h, and then allowed to come to r.t.over 1 h. The mixture is diluted with ether, washed with brine, dried(MgSO₄), and evaporated. HPLC purification (hexane:EtOAc 88:12) gives12.15 (110 mg, 85%).

Rf: 0.41 (hexane:EtOAc 8:2).

IR (film): 1708, 1639, 1616, 1004 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 7.23 (1H, dd, J=15.39, 11.00); 6.12 (1H,dd, J=15.19, 10.99); 5.93 (1H, dd, J=15.19, 9.66); 5.80 (1H, d,J=15.39); 4.20 (2H, m); 1.29 (3H, dd, J=7.08, 7.08); 0.96 (3H, d,J=6.65); 1.01 (3H, s) ppm.

EXAMPLE 76

Synthesis of 12.14

A mixture of 12.13 (67 mg, 0.23 mmol), 5% Rh/Al₂O₃ (30 mg) and EtOAc (4ml) is stirred under H₂ (atmospheric pressure) at r.t. for 1.5 h. Themixture is then filtered through a short silica gel column (hexane:EtOAc1:1). HPLC purification (hexane:EtOAc 88:12) gives 12.14 (63 mg, 93%).

Rf: 0.49 (hexane:EtOAc 8:2).

IR (film): 1735, 1707 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.12 (2H, q, J=7.20 Hz); 1.25 (3H, t,J=7.20 Hz), 1.02 (3H, s); 0.82 (3H, d, J=6.65) ppm.

EXAMPLE 77

Synthesis of 12.15

The Horner-Wittig coupling of 12.14 (60 mg, 0.19 mmol) with 13.2 (162mg, 0.28 mmol), using n-BuLi (1.6 M solution in hexane, 175 μl, 0.28mmol) as base, is carried out as described for 10. Flash chromatography(hexane:EtOAc 1:1) and HPLC separation (hexane:EtOAc 18:1) gives (80 mg,62%).

Rf: 0.64 (hexane:EtOAc 9:1).

EXAMPLE 78

Synthesis of 12.17

From 12.8 by Horner-Wittig reaction as described for 12.15 from 12.9followed by NaOEt-EtOH induced epimerization at r.t. for 21 h (overallyield 48%).

Rf: 0.28 (n.pentane:acetone 94:6).

IR (film): 2957 (s); 1713 (s); 1641 (s); 1463 (m); 1137 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 7.21 (1H, dd, J=10.8, 15.3); 6.10 (1H, dd,J=10.8. 15.1); 5.99 (1H, dd, J=8.8, 15.1); 5.77 (1H, d, J=15.3); 4.19(2H, q, J=7.1); 2.32 (4H, m); 2.15(1H, m); 1.92 (1H, m); 1.84 (1H, m);1.75 (3H, m); 1.60 (2H, m); 1.44 (1H, m); 1.35 (1H, m); 1.29 (3H, t,J=7.1); 1.05 (3H, d, J=6.5); 1.04 (3H, s) ppm.

EXAMPLE 79

Synthesis of 12.18

From 12.17 as described for 12.16 from 12.15 (yield: 88%).

Rf: 0.35 (n.pentane:acetone 96:4).

IR (film): 2954 (s); 1733 (s); 1713 (s); 1380 (s); 1159 (s); 1097 (s)cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.12 (2H, q, J=7.1); 2.31 (5H, m); 2.15(1H, m); 1.91 (3H, m); 1.75 (2H, m); 1.56 (3H, m); 1.43-1.24 (6H, m);1.25 (3H, t, J=7.1); 1.19 (1H, m); 1.03 (3H, s); 0.89 (3H, d, J=6.6)ppm.

EXAMPLE 80

Synthesis of Analogue 1

As described for 11 starting from 13.1 and 10-hydroxy-10-methyldecanal.

Rf: 0.40 (dichloromethane:methanol 9:1).

IR (film): 3374 (s); 3025 (w); 2969 (s); 2929 (s); 2853 (s); 1634 (m);1466 (w); 1432 (w); 1366 (w); 1306 (w); 1266 (w); 1218 (w); 1149 (w);1054 (w); 975 (w); 958 (w); 907 (w); 800 (w); 737 (w) cm⁻¹.

¹ H NMR: (360 MHz, CDCl₃): δ: 6.37 (1H, dd, J=11, 15 Hz); 6.03 (1H, d,J=11 Hz); 5.71 (1H, dt, J=15 Hz); 5.31 (1H, d, J=7 Hz); 4.99 (1H, d);4.42 (1H, t, J=5.5 Hz); 4.20 (1H, m); 2.58 (1H, dd, J=13 Hz); 2.54 (1H,dd, J=4 Hz°; 2.06 (2H, dd, J=7 Hz); 1.96 (2H, t, J=5.51 Hz); 1.85-1.65(3H, m); 1.50-1.15 (18H, m).

EXAMPLE 81

Synthesis of the Analogue 2

From 1.8 d as described for 13.

Rf: 0.37 (dichloromethane:methanol 88:12).

IR (film): 3368; 1610; 1374; 1049 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.31 (1H, d, J=11.2); 6.06 (1H, d, J=11.2);4.15 (1H, bs); 4.13-4.04 (2H, m); 2.75-2.64 (2H, m); 2.49 (1H, dd,J=13.1, 3.8); 2.40 (1H, m); 2.28 (1H, dd, J=13.8, 7.9); 2.21 (1H, dd,J=13.5, 7.1); 2.15-0.70 (22H, ); 1.21 (6H, s); 0.9 (3H, d, J=6.73) ppm.

EXAMPLE 82

Synthesis of the Analogue 3

From 1.11d as described for 11.

Rf: 0.32 (acetone:hexane 4:6).

IR (film) 3356; 1441; 1378; 1215; 1144 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.37 (1H, d, J=11.4); 6.18 (1H, d, J=11.4);5.32 (1H, bs); 5.02 (1H, s); 4.43 (1H, m); 4.21 (1H, m); 2.87 (1H, dm,J=13.6); 2.61 (1H, dd, J=3.4 Hz, J=13.2 Hz); 2.30 (1H, dd, J=7.3, 13.2);2.01-1.93 (2H, m); 1.25 (6H, s, 20% 20-epi); 1.21 (6H, s); 0.87 (3H, d,J=6.7); 0.76 (3H, d, J=6.6, 20% 20-epi) ppm.

EXAMPLE 83

Synthesis of the Analogue 4

To a solution of 13.1 (76 mg, 0.13 mmol) in THF (2 ml) was addeddropwise n-butyllithium (52 μl, 0.13 mmol, 2.5M solution in hexane) at−78° C. under nitrogen atmosphere. The formed dark red solution wasstirred for 1 hour at −78° C. after which a solution of 2.5 (25 mg,0.065 mmol) in THF (1 ml) was added. The red solution was stirred at−78° C. for 1 hour and was then warmed up to room temperature. Thereaction mixture was immediately filtered through a silica gel column(EtOAc:Hex 1:30) and the crude product (74 mg) was further purified byHPLC (EtOAc:Hex 1:200) yielding 45.0 mg (92%) of coupling product.

A solution of coupling product (45.0 mg, 0.06 mmol) and TBAF (1.27 ml,1.27 mmol, 1M solution in THF) in THF (3 ml) was stirred at roomtemperature (25-30° C.) for 39 hours. The reaction mixture wasimmediately filtered through a silica gel column (MeOH:CH₂Cl₂ 1:20) andthe crude product (59 mg) was separated by HPLC (MeOH:CH₂Cl₂ 1:16) togive 4 (19.1 mg, 78%). A product (8.3 mg), that was not identified, wasalso obtained.

Rf: 0.21 (dichloromethane:methanol 1:20).

IR (film): 3378 (s); 2954 (s); 1643 (w); 1453, 1383 (s); 1264 (s); 1142(w); 1057 (s); 742 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.32 (1H, d, J=11.2 Hz); 6.05 (1H, d,J=11.2 Hz); 5.32 (1H, m); 4.98 (1H, m); 4.45 (1H, m); 4.20 (1H, m); 3.30(1H, m); 2.60 (1H, dd, J=3.9, 13.2 Hz); 2.42 (1H, m); 2.30 (1H, dd,J=7.4, 13.2 Hz); 2.24 (2H, m); 1.96 (2H, m); 1.79 (1H, d, J=13.1 Hz);1.65 (6H, m); 1.47 (2H, m); 1.25 (1H, m); 0.90 (3H, d, J=6.6 Hz); 0.89(6H, dd, J=6.8 Hz); 0.75 (3H, s); 0.74 (3H, d, J=7.2 Hz).

EXAMPLE 84

Synthesis of analogue 5 and Previtamin 56

To a solution of 13.1 (110 mg, 0.188 mmol) in THF (3 ml) was addeddropwise n-butyllithium (76 μl, 0.188 mmol, 2.5M solution in hexane) at−78° C. under nitrogen atmosphere. The formed dark red solution wasstirred for 1 hour at −78° C. after which a solution of the 2.7 (36 mg,0.094 mmol) in THF (1 ml) was added. The red solution was stirred at−78° C. for 1 hour and was then slowly warmed up to room temperature.The reaction mixture was immediately filtered through a silica gelcolumn (EtOAc:Hex 1:20) and the crude product (117 mg) was furtherpurified by HPLC (EtOAc:Hex 1:200) yielding 66.0 mg (93%) of couplingproduct.

A solution of coupling product (65.0 mg, 0.087 mmol) and TBAF (2.61 ml,2.61 mmol, 1M solution in THF) in THF (8 ml) was stirred at 30-40° C.for 40 hours. The reaction mixture was immediately filtered through asilica gel column (MeOH:CH₂Cl₂ 1:20) and the crude product (82 mg) wasseparated again by HPLC (MeOH:CH₂Cl₂ 1:20) to give 5 (23.3 mg, 66%) and56 (4.2 mg, 12%).

5: Rf: 0.15 (dichloromethane:methanol 1:20).

IR (film): 3385 (s); 2956 (s); 1642 (w); 1450, 1383 (s); 1056, 909 (s);734 (m) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.32 (1H, d, J=11.2 Hz); 6.06 (1H, d,J=11.2 Hz); 5.31 (1H, d, J=2.2 Hz); 4.99 (1H, d, J=2.2 Hz); 4.42 (1H,m); 4.21 (1H, m); 3.31 (1H, m); 2.60 (1H, dd, J=3.9, 13.2 Hz); 2.42 (1H,m); 2.29 (1H, dd, J=7.4, 13.1 Hz), 2.02 (4H, m); 1.80 (1H, d, J=13.0Hz); 1.65 (5H, m); 1.45 (4H, m); 0.90 (3H, d, J=6.6 Hz); 0.89 (6H, dd,J=6.6 Hz); 0.75 (3H, s); 0.74 (3H, d, J=7.2 Hz).

56: Rf: 0.15 (dichloromethane:methanol 20:1).

IR (film): 3384 (s), 2956, 2863 (s), 1640 (w), 1453, 1383 (s), 1056, 908(s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 5.89 (1H, d, J=12.2 Hz); 5.68 (1H, d,J=13.0 Hz); 5.65 (1H, br s); 4.18 (1H, br s); 4.10 (1H, m); 3.33 (1H,m); 2.43 (1H, dd, J=4.5, 16.8 Hz); 2.10 (2H, m); 2.00 (2H, m); 1.72 (3H,s); 1.71 (2H, m); 1.60 (4H, m); 1.45 (3H, m); 1.33 (2H, m); 0.92 (3H, d,J=6.8 Hz); 0.90 (3H, d, J=6.6 Hz); 0.88 (3H, d, J=5.9 Hz); 0.80 (3H, s);0.77 (3H, d, J=7.2 Hz).

MS (m/z): 404 (5), 387 (6), 386 (4), 229 (30), 211 (30), 95 (40).

EXAMPLE 85

Synthesis of 6

To a solution of 13.2 (57 mg, 0.099 mmol) in THF (2 ml) was addeddropwise n-butyllithium (40 μl, 0.099 mmol, 2.48M solution in hexane) at−78° C. under nitrogen atmosphere. The formed dark red solution wasstirred for 1 hour at −78° C. after which a solution of 2.7 (19 mg,0.049 mmol) in THF (1.5 ml) was added. The red solution was stirred at−78° C. for 1 hour and was then slowly warmed up to room temperature.The reaction mixture was immediately filtered through a silica gelcolumn (EtOAc:Hex 1:20) and the crude product 44 mg) was furtherpurified by HPLC (EtOAc:Hex 1:140) yielding 32.0 mg (88%) of couplingproduct.

A solution of coupling product (32.0 mg, 0.043 mmol) and TBAF (1.96 ml,1.96 mmol, 1M solution in THF) in THF (4 ml) was stirred at 30-45° C.for 40 hours. The reaction mixture was immediately filtered through asilica gel column (MeOH:CH₂Cl₂ 1:20) and the crude product (17 mg) wasseparated again by HPLC (MeOH:CH₂Cl₂ 1:20) to give a 4/1 mixture of E-and Z-isomers (15.5 mg, 91%).

This mixture was separated again on a special HPLC column (RSiL CN, 10micron; 5.0 ml/min; 5.0 mg/500 μl/shot) with eluent Hex:i.PrOH:CH₃C N89:10:1 to give 13.2 mg of analogue 6 (E-isomer) and 2 mg of Z-isomer.The separation was not easy and both were separated several times.

IR (film): 3380 (s); 2957 (s); 1616 (w); 1452, 1381 (m); 1047 (s); 736(m) cm⁻¹.

¹H NMR: (CDCl3): δ6.26 (1H, d, J=11.2); 5.94 (1H, d, J=11.2); 4.08 (2H,m); 3.33 (1H, m); 2.64 (1H, dd, J=3.8, 13.3); 2.48 (1H, dd, J=3.7,13.3); 2.43 (1H, m); 2.29 (1H, dd, J=7.7, 13.4); 2.18 (1H, dd, J=6.7,13.3); 2.07 (1H, d, J=13.0); 2.00 (1H, m); 1.88 (2H, m); 1.86 (1H, d,J=13.1); 0.93 (3H, d, J=6.3); 0.91 (3H, d, J=6.4); 0.89 (3H, d, J=6.5);0.79 (3H, d, J=7.8); 0.87 (3H, s) ppm.

MS (m/z): 392 (M.⁺, 5); 374 (8); 308 (10); 235 (50); 217 (40); 55 (100).

EXAMPLE 86

Synthesis of Analogue 7

As described for 11.

IR (film): 3380 (s); 2939 (s); 1625 (w); 1452, 1383 (m); 909 (m) cm⁻¹.

¹H NMR: (CDCl3): δ66.32 (1H, d, J=11.2); 6.04 (1H, d, J=11.2); 5.32 (1H,t, J=1.4); 4.99 (1H, m); 4.43 (1H, m); 4.22 (1H, m); 2.60 (1H, dd,J=3.9, 13.2); 2.43 (1H, dt, J=13.7, 5.1); 2.29 (1H, dd, J=7.4,13.2);2.03 (1H, m); 2.02 (1H, d, J=13.1); 1.96 (2H, m); 1;80 (1H, d,J=13.0); 1.21 (6H, s); 0.89 (3H, d, J=6.8); 0.75 (3H, s); 0.73 (3H, d,J=6.3) ppm.

MS (m/z): 404 (M.⁺, 1%).

EXAMPLE 87

Synthesis of Analogue 8

As described for 13.

UV: λ_(max)=249.5 nm.

IR (film): 3382 (s); 2935 (s); 1615 (w); 1454, 1380 (m); 1048 (m); 909,734 (s) cm⁻¹.

¹H NMR: (CDCl3): δ6.26 (1H, d, J=11.2); 5.94 (1H, d, J=11.2); 4.09 (2H,m); 2.67 (1H, dd, J=3.8, 13.3); 2.49 (1H, dd, J=3.8, 13.3); 2.43 (1H,m); 2.29 (1H, dd, J=7.6, 13.3); 2.19 (1H, dd, J=6.7, 13.2); 2.04 (1H, d,J=13.0 Hz); 2.00 (1H, m); 1.90 (1H, m); 1.86 (1H, m); 1.84 (1H, d,J=13.0 Hz); 1.70 (1H, m); 1.56 (3H, m); 1.21 (6H, s); 0.89 (3H, d,J=6.9); 0.77 (3H, d, J=7.3); 0.76 (3H, s) ppm.

EXAMPLE 88

Synthesis of Analogue 9

As described for 11.

Rf: 0.30 (dichloromethane:methanol 1:20).

IR (film): 3386 (s); 2932, 2874 (s); 1640 (w); 1456, 1475 (s); 1141,1053 (s); 816 (m) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.32 (1H, d, J=11.3 Hz); 6.10 (1H, d,J=11.3 Hz); 5.25 (1H, d, J=1.7 Hz); 5.05 (1H, d, J=2.2 Hz); 4.40 (1H,m); 4.24 (1H, m); 3.65 (2H, m); 3.46 (2H, m); 2.62 (1H, dd, J=4.1, 12.8Hz); 2.47 (1H, m); 2.25 (1H, dd, J=10.8, 12.3 Hz); 2.12 (2H, m); 2.02(1H, m); 1.80 (3H, m); 1.70 (3H, m); 1.55 (4H, m); 1.35 (2H, m); 1.23(6H, s); 0.8 (3H, s).

EXAMPLE 89

Synthesis of Analogue 10

As described for 11. Both epimers could be separated by HPLC(silica:ethyl acetate:pentane 15:85) on the stage of the TBMBS ethers.The respective structures were proven bij NOE measurements.

Rf: 0.36 (dichloromethane:methanol 7:1).

10α: Rf: 0.20 dichloromethane:methanol 92:8).

IR (film): 3324, 2986, 2880, 1455, 1414, 1378, 1260, 1130 cm⁻¹.

¹H NMR: (MHz, CDCl3): δ: 6.45 (1H, dd, J=11, 15 Hz); 6.05 (1H, d, J=11Hz); 5.71 (1H, dt, J=7.5 and 15 Hz); 5.30 (1H, m); 5.00 (1H, d, J=4.5Hz); 4.98 (1H, m); 4.42 (1H, m); 4.21 (1H, m); 3.80 (1H, d, J=8.2 Hz);3.50 (1H, d, J=8.2 Hz); 3.48 (1H, s); 2.58 (1H, dd, J=4, 13.2 Hz); 2.35(2H, d, J=7.5 Hz); 2.28 (1H, dd, J=7.2,13.2 Hz); 1.95 (2H, t, J=5.5 Hz);1.7-1.5 (4H, m); 1.48 (4H, q, J=7.5 Hz); 1.43 (2H m); 1.25 (3H, s); 0.85(6H, t, J=7.5 Hz).

10β: ¹H NMR: (500 MHz, CDCl3): δ: 6.46 (1H, dd, J=11, 15 Hz); 6.05 (1H,d, J=11 Hz); 5.69 (1H, dt, J=7.5,15 Hz); 5.31 (1H, t, J=1.5 Hz); 5.00(1H, t, J=4.7 Hz); 4.98 (1H, m); 4.45 (1H, m); 4.21 (1H, m); 3.67 (1H,d, J=8 Hz); 3.62 (1H, d, J=8 Hz); 2.58 (1H, dd, J=3.4, 13.3 Hz); 2.40(1H, dd, J=7.5, 14 Hz); 2.35 (1H, dd, J=7.5, 14 Hz); 2.28 (1H, dd, J=7,13.3 Hz); 1.97 (2H, t, J=5.5 Hz); 1.75-1.55 (4H, m); 1.47 (4H, q, J=7.5Hz); 1.43 (2H, m); 1.27 (3H, s); 0.85 (6H, t, J=7.5 Hz).

EXAMPLE 90

Synthesis of Analogue 11

To a solution of dry A-ring phosphine oxide 13.1 (87 mg, 150 μmol) intetrahydrofuran (1.4 ml) is added a n.butyllithium solution (2.5 M inhexane, 57 μl, 142.5 μmol) at −78° C. After stirring, the resulting redsuspension for 1 hour, a solution of 6.12 (12 mg, 47.2 μmol) intetrahydrofuran (0.5 ml) is added dropwise. The reaction mixture isstirred for 1 hour at −78° C. and then the cooling bath is removed.Water is added slowly till the orange colour has completely disappearedand the tetrahydrofuran is removed. After addition of diethylether andsaturated sodium bicarbonate, the aqueous layer is extracted severaltimes with diethylether. The collected organic phases are filteredthrough silicagel, the filtrate concentrated in vacuo and the remainingoil purified by HPLC (pentane:ethyl acetate 8:2) to give 24 mg (82%) ofthe coupled product.

To a solution of this (24 mg, 38.8 μmol) in tetrahydrofuran (0.5 ml) isadded a tetra (n.butyl)ammoniumfluoride solution (1 M in THF, 311 μl,311 μmol) and the resulting mixture is stirred at room temperature underan argon atmosphere in the dark for 87 hours. After evaporation of theTHF, the residue is purified on a silica column(dichloromethane:methanol 9:1) and HPLC (CH₂Cl₂:MeOH 94:6) to yield 15mg (98%) of 11.

Rf: 0.16 (dichloromethane:methanol 9:1).

IR (film): 3354 (s); 2966 (s); 2936 (o); 2869 (m); 1635 (w); 1468 (m);1377 (s); 1366 (s); 1265 (m); 1215 (m); 1152 (s); 1056 (s); 976 (m)cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 6.36 (1H, dd, J=11 Hz, J=15 Hz); 6.15 (1H,d, J=11 Hz); 5.77 (1H, dt, J=15 Hz, J=7.5 Hz); 5.31 (1H, d, J<1); 5.00(1H, d); 4.43 (1H, t, J=5.5); 4.21 (1H, m); 2.57 (1H, dd, J=13, 3.7 Hz);2.27 (1H, dd, J=7); 2.10-1.71 (7H, m); 1.60-1.25 (10 H, m); 1.21 (6H,s); 0.82 (3H, s); 0.78 (3H, s); 0.66 (3H, s).

EXAMPLE 91

Synthesis of Analogue 12

As described for 11.

Rf: 0.25 (dichloromethane:methanol 95:5).

IR (film): 3374 (s, br); 2965 (s); 2938 (s); 2876 (m); 1631 (m); 1460(m); 1057 (s); 976 (m); 935 (s); 909 (s) cm⁻¹.

¹H NMR: (200 MHz, CDCl₃): δ: 6.36 (1H, dd, J 10.7, 15 Hz); 6.05 (1H, d,J=10.7 Hz); 5.76 (1H, dt, J=15, 7.5 Hz); 5.31 (1H, d); 4.99 (1H, d);4.43 (1H, t, J=5.6 Hz); 4.21 (1H, m); 2.58 (1H, dd, J=7, 13.2 Hz); 2.25(1H, dd, J=3.7, 13.2 Hz); 2.06-2.00 (1H, m); 2.00-1.92 (2H, t);1.84-1.54 (5H, m); 1.53-1.38 (6H, m); 1.38-1.00 (8H, m); 0.87 (6H, t);0.83 (3H, s); 0.79 (3H, s); 0.66 (3H, s).

EXAMPLE 92

Synthesis of Analogue 13

To a solution of 13.2 (76 mg, 133 μmol) in tetrahydrofuran (1.3 ml) isadded a n.butyllithium solution (2.5M in hexane, 51 μl, 127 μmol) at−75° C. After stirring the resulting red suspension during 1 h, asolution of 6.13 (13 mg, 46 μmol) in tetrahydrofuran (0.5 ml) is addeddropwise. The reaction mixture is stirred for 1 h at −75° C. andsubsequently the cooling bath is removed. After addition of ethylacetate (2 ml) and saturated sodium bicarbonate (2 ml) the aqueous layeris extracted several times with ethyl acetate. The collected organicphases are filtered through silicagel, the filtrate concentrated invacuo and the remaining oil purified by HPLC (pentane:ethyl acetate95:5) to give 11 mg (38%) of the coupled product.

To a solution of this (11 mg, 17 μmol) in methanol (2.5 ml) andtetrahydrofuran (2.5 ml) is added Amberlyst-15 (1.6 9) and the resultingmixture is stirred at room temperature under argon in the dark for 9 h.The mixture is filtered through silicagel. The Amberlyst-15 is washedseveral times with methanol and filtered through silicagel. The filtrateis concentrated in vacuo and the remaining oil purified by HPLC(dichloromethane:methanol 95:5) to give 13 (6 mg, 86%).

Rf: 0.19 (dichloromethane:methanol 95:5).

UV: λ_(max)240,9 nm; (ε=32.535,7).

IR (film): 3443 (s, br); 2913 (w), 1518 (m); 1433 (s); 1256 (w); 1087(w), 1024 (w) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.26 (1H, dd, J=10.8, 14.9 Hz); 6.05 (1H,d, J=10.8 Hz); 5.72 (1H, dt, J=15, 7.7 Hz); 4.12-4.08 (2H, m); 2.54 (1H,dd, J=13.5, 3.9 Hz); 2.47 (1H, dd, J=13.1, 3.5 Hz); 2.3 (1H, dd, J=13.3,7.6 Hz); 2.15 (1H, dd, J=13.3, 6.9 Hz); 2.1 (1H, dd, J=13.3, 7.1 Hz);2.03 (1H, dd, J=13.6, 8.4 Hz); 1.8-1.9 (3H, m); 1.75 (1H, m); 1.64-1.5(3H, m); 1.48-1.4 (4+2, q+m); 1.4-1 (7H, m); 0;87 (6H, t, J=7.4 Hz);0.84 (3H, s); 0.79 (3H, s); 0.67 (3H, s).

EXAMPLE 93

Synthesis of Analogue 14

The coupling of 6.21α with 13.1 is carried out as described for 6.12.Cleavage of the silyl ether is however performed upon stirring amethanolic solution in the presence of Amberlyst 15 for 4 h at roomtemperature. After filtration the compound 14 is purified by HPLC(dichloromethane:methanol 95:5).

Rf: 0.40 (dichloromethane:methanol 9:1).

IR (film): 3386 (s); 2969 (s); 1641 (w); 1468 (m); 1366 (s); 1154 (m);1058 (s); 978 (m) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.37 (1H, dd, J=10.8, 15.1 Hz); 6.04 (1H,d, J=10.8 Hz); 5.71 (1H, dt, J=15.1 Hz, J=7.5 Hz); 5.30 (1H, d, J<1);4.98 (1H, d); 4.42 (1H, m); 4.21 (1H, m); 3.75 (1H, dt); 3.61-3.55 (2H,m); 2.56 (1H, dd, J=13.2, 3.9 Hz); 2.25 (1H, dd, J=7.3 Hz); 2.11-1.90(4H, m); 1.81-1.30 (9H, m); 1.24 (6H, s); 0.89 (3H, s); 0.83 (3H, s);0.81 (3H, s).

EXAMPLE 94

Synthesis of Analogue 15

As described for 14 starting from 6.22α.

Rf: 0.59 (dichloromethane:methanol 9:1).

IR (film): 3380 (s, br); 2964 (s); 1632 (w); 1462 (s); 1376 (s); 1262(m); 1067 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.37 (1H, dd, J=10.8, 15.1 Hz); 6.04 (1H,d); 5.71 (1H, dt, J=7.5 Hz); 5.30 (1H, d); 4.98 (1H, d); 4.42 (1H, m);4.21 (1H, m); 3.71 (1H, m); 3.60-3.46 (2H, m); 2.57 (1H, dd, J=13.4, 3.7Hz); 2.25 (1H, dd, J=7.4 Hz); 2.11-1.9 (4H, m); 1.73-1.65 (3H, m);1.60-1.20 (10H, m); 0.89 (3H, s); 0.87 (6H, t); 0.83 (3H, s); 0.81 (3H,s).

EXAMPLE 95

Synthesis of Analogue 16

As described for 14 starting from 6.16.

Rf: 0.27 (dichloromethane:methanol 95:5).

IR (film): 3380 (s, br); 2966 (s); 1616 (w); 1551 (m); 1422 (s); 1278(s); 1156 (m) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 6.35 (1H, dd, J=10.9, 15.2 Hz); 6.07 (1H,d, J=10.9 Hz); 5.76 (1H, dt); 5.31 (1H, d, J<1); 5.00 (1H, d); 4.44 (1H,t, J=5.8 Hz); 4.22 (1H, m); 2.57 (1H, dd, J=13.3, 3.6 Hz); 2.27 (1H, dd,J=6.8 Hz); 2.10-1.50 (11H, m); 1.45 (4H, q, J=7.5 Hz); 1.42-1.00 (7H,m); 0.86 (6H, t, J=7.5 Hz); 0.80 (3H, s); 0.78 (3H, s); 0.67 (3H, s).

EXAMPLE 96

Synthesis of Analogue 17

As described for 11.

Rf: 0.31 (dichloromethane:methanol 9:1).

IR (film): 3384 (s); 2968 (s); 1631 (m); 1467 (s); 1365 (s); 1265 (m);1154 (m); 1090 (s); 1056 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.35 (1H, dd, J=10.6, 15 Hz); 6.05 (1H, d,J=10.6); 5.73 (1H, dt, J=15, 7.5 Hz); 5.31 (1H, d); 4.99 (1H, d, J˜1);4.43 (1H, t, J=5.5 Hz); 4.22 (1H, m); 3.75 (1H, m); 3.55 (1H, m) 3.48(1H, m); 2.57 (1H, dd); 2.27 (1H, dd); 2.00 (4H, m); 1.75 (2H, t, J=5.6Hz); 1.83-1.20 (7H, m); 1.25 (6H, s); 0.88 (3H, s); 0.86 (3H, s); 0.83(3H, s).

EXAMPLE 97

Synthesis of Analogue 18

As described for 14 starling from 6.22β.

Rf: 0.54 (dichloromethane:methanol 9:1).

IR (film): 3380 (s, br); 2964 (s); 2875 (s); 1632 (w); 1462 (s); 1364(m); 266 (m); 1091 (s); 1065 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.35 (1H, dd, J=10.7, 15.1 Hz); 6.05 (1H,d, J=10.7 Hz); 5.72 (1H, dt, J=7.5 Hz); 5.31 (1H, d); 4.99 (1H, d); 4.43(1H, m); 4.22 (1H, m); 3.71 (1H, dt); 3.55-3.45 (2H, m); 2.57 (1H, dd,J=13.3, 3.7 Hz); 2.28 (1H, dd, J=7.0 Hz); 2.02-1.90 (4H, m); 1.72 (2H,t); 1.61-1.43 (11H, m); 0.87 (3H, s); 0.85 (3H, s); 0.84 (6H, t); 0.83(3H, s).

EXAMPLE 98

Synthesis of the Analogue 19

To a solution of 6.27 (12 mg, 19 μmol) in THF (1 ml), at −5° C., asolution of MeMgBr (50 μl 3M in Et₂O, 8 eq) is added dropwise. Afterwarming overnight to r.t. the mixture is poured in anice-ammoniumchloride solution-ether mixture. The organic phase is dried(MgSO₄). Filtration, evaporation and column chromatography (diethylether:hexane 1:9→1:4) gives the bis silylated analogue (10 mg, 82%).TBAF deprotection as described for analogue 11 gives 19 (5 mg, 80%).

Rf: 0.27 (MeOH:CH₂Cl₂ 1:19).

¹H NMR: (500 MHz, CDCl₃): δ: 6.36 (1H, dd, J=10.8, 15.2); 6.05 (1H,10.8); 5.76 (1H, dt, J=7.7, 15.1); 5.31 (1H, dd, J=1,2); 5.00 (1H, brs); 4.43 (1H, t, J 5.7); 4.22 (1H, m); 2.57 (1H, dd, J=3.8, 13.3); 2.26(1H, dd, J=7.5, 13.3); 1.21 (6H, s); 0.82 (3H, s); 0.78 (3H, s); 0.66(3H, s) ppm.

EXAMPLE 99

Synthesis of the Analogue 20

From 6.27 with EtMgBr as described for 19 (yield 50%).

Rf: 0.29 (MeOH:CH₂Cl₂ 1:19).

¹H NMR: (500 MHz, CDCl₃): δ: 6.36 (1H, dd, J=10.8, 15.1); 6.06 (1H,d=10.8); 5.76 (1H, dt, J=7.5, 15.1); 5.31 (1H, dd, J=1, 2); 5.00 (1H, brs); 4.43 (1H, t, J=5.5); 4.22 (1H, m); 2.57 (1H, dd, J=3.6, 13.2); 2.26(1H, dd, J=7.2, 13.3); 1.46 (4H, q, 7.5); 0.86 (6H, t, J=7.5); 0.82 (3H,s); 0.78 (3H, s); 0.66 (3H, s) ppm.

EXAMPLE 100

Synthesis of 21

From 6.30 as described for 19 from 6.26 (yield 48%).

Rf: 0.16CH₂Cl₂: (MeOH 1:19).

¹H NMR: (500 MHz, CDCl₃): δ: 6.36 (1H, dd, J=10.8, 15.1); 6.06 (1H,d=10.8); 5.76 (1H, dt, J=7.5, 15.1); 5.31 (1H, dd, J=1, 2); 5.00 (1H, brs); 4.43 (1H, t, J=5.5); 4.22 (1H, m); 2.57 (1H, dd, J=3.6, 13.2); 2.26(1H, dd, J=7.2, 13.3); 1.46 (4H, q, 7.5); 0.86 (6H, t, J=7.5); 0.82 (3H,s); 0.78 (3H, s); 0.66 (3H, s) ppm.

EXAMPLE 101

Synthesis of Analogue 22

As described for 11.

Rf: 0.30 (dichloromethane:methanol 1:20).

IR (film): 3389 (s); 2932 (s); 1632 (w); 1462, 1366 (m); 1089, 1057 (s);736 (m) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.38 (1H, dd, J=10.7, 15.1 Hz); 6.04 (1H,d, J=10.7 Hz); 5.72 (1H, m); 5.30 (1H, s); 4.98 (1H, s); 4.41 (1H, m);4.21 (1H, m); 3.73 (1H, m); 3.66 (1H, m); 3.38 (1H, m); 2.58 (1H, m);2.27 (1H, m); 1.97 (4H, m); 1.73 (2H, m); 1.60 (4H, m); 1.40 (2H, m);1.22 (6H, s); 0.87 (3H, s).

EXAMPLE 102

Synthesis of Analogue 23

As described for 11.

Rf: 0.21 (dichloromethane:methanol 1:17).

IR (film): 3384, 2932 (s); 1630 (w); 1455, 1365 (m); 1265, 1152 (m);1089, 1054 (s); 909, 734 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.38 (1H, m); 6.05 (1H, d, J=10.8 Hz); 5.70(1H, m); 5.28 (1H, s); 4.95 (1H, s); 4.41 (1H, m); 4.21 (1H, m); 3.70(2H, m); 3.41 (1H, m); 2.55 (1H, m); 2.25 (1H, m); 2.15 (2H, m); 2.00(4H, m); 1.72 (4H, m); 1.60 (2H, m); 1.40 (2H, m); 1.21 (6H, s); 1.10(2H, m); 0.89 (3H, s).

EXAMPLE 103

Synthesis of Analogue 24

As described for 11.

Rf: 0.29 (dichloromethane:methanol 1:20).

IR (film): 3421 (m); 2931 (s); 1637 (w); 1458, 1379 (m); 1085 (s); 911(s); 935 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.38 (1H, m); 6.04 (1H, d, J=10.8 Hz); 5.70(1H, m); 5.30 (1H, m); 4.98 (1H, s); 4.40 (1H, m); 4.20 (1H, m); 3.58(1H, m); 3.53 (1H, m); 3.43 (1H, m); 2.92 (1H, m); 2.55 (1H, m); 2.25(1H, m); 2.08 (1H, m); 1.98 (4H, m); 1.80 (3H, m); 1.60 (1H, m); 1.40(3H, m); 1.32 (3H, s); 1.28 (3H, s); 0.90 (3H, s).

EXAMPLE 104

Synthesis of Analogue 25

As described for 11.

Rf: 0.30 (dichloromethane:methanol 1:20).

IR (film): 3401 (s); 2924 (s); 1633 (w); 1453, 1374 (m); 1164, 1054 (s);738 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.39 (1H, m); 6.06 (1H, d, J=10.8 Hz); 5.70(1H, m); 5.32 (1H, t, J=1.6 Hz); 5.00 (1H, m); 4.43 (1H, m); 4.21 (1H,m); 2.55 (1H, m); 2.25 (1H, m); 2.15 (1H, dd, J=8.2, 14.0 Hz); 2.00 (5H,m); 1.90 (2H, m); 1.60 (4H, m); 1.49 (6H, s); 1.40 (2H, m); 1.10 (1H,m); 0.87 (3H, s).

EXAMPLE 105

Synthesis of Analogue 26

As described for 11.

Rf: 0.36 (dichloromethane:methanol 7:1).

¹H NMR: (500 MHz, CDCl₃): δ: 6.39 (1H, dd, 10.8, 15.2 Hz); 6.04 (1H, d,10.8 Hz); 5.68 (1H, dt, 7, 15 Hz); 5.31 (1H, dd, 1, 2 Hz); 4.99 (1H, d,1 Hz); 4.43 (1H, m); 4.22 (1H, m); 3.72 (1H, ddd, 5, 7, 9.5 Hz); 3.66(1H, ddd, 5, 7, 9.5 Hz); 3.27 (1H, dt, 4, 11 Hz); 2.57 (1H, dd, 3.7,13.3 Hz); 2.27 (1H, dd, 7.0, 13.4 Hz); 1.24 (6H, s); 0.76 (3H, d, 7.02Hz).

EXAMPLE 106

Synthesis of Analogue 27

As described for 11.

Rf: 0.23 (dichloromethane:methanol 9:1).

IR (film): 3382 (s); 2930 (s); 1632, 1445, 1359, 1261, 1153, 1091 cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 6.37 (1H, dd, 10.8, 15.1 Hz); 6.03 (1H, d,10.8 Hz); 5.66 (1H, dt, 7.5, 15.1 Hz); 5.30 (1H, br s); 4.98 (1H, br s);4.43 (1H, m); 4.21 (1H, m); 3.87 (1H, ddd, 4.5, 7, 9 Hz); 3.53 (1H, ddd,5, 7, 9 Hz); 2.90 (1H, m); 2.80 (1H, td, 4, 10 Hz); 2.57 (1H, dd, 3.6,13.3 Hz); 2.31 (1H, m); 2.25 (1H, dd, 7.4, 13.3 Hz); 2.10 (1H, m); 1.24(6H, s); 1.01 (3H, d, 6.03 Hz).

EXAMPLE 107

Synthesis of Analogue 28

As described for 11.

Rf: 0.26 (dichloromethane:methanol 9:1).

IR (film): 3384 (s); 2929 (s); 3026, 1631, 1443, 1363, 1261, 1218 cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 6.35 (1H, dd, 10.8, 15.2 Hz); 6.02 (1H, d,10.8 Hz); 5.67 (1H, dt, 7, 15 Hz); 5.29 (1H, d, 1 Hz), 4.97 (1H, d, 1Hz), 4.42 (1H, m); 4.21 (1H, m); 3.84 (1H, ddd, 4, 7, 9 Hz); 3.49 (1H,ddd, 4, 7, 9 Hz); 3.36 (1H, W1/2, 8 Hz, m); 2.56 (1H, dd, 4,13 Hz); 2.25(1H, dd, 7, 13 Hz); 2.19 (1H, m); 1.25 (6H, s); 0.98 (3H, d, 6.7 Hz).

EXAMPLE 108

Synthesis of Analogue 29

As described for 11.

Rf: 0.38 (dichloromethane:methanol 9:1).

IR (film): 3357, 2926, 2857, 1366, 1056, 975, 908, 801, 734 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.34 (1H, dd, J=15.2, 11.0 Hz); 6.04 (1H,d, J=10.9 Hz); 5.67 (1H, ddd, J=15.1, 8.5, 6.0 Hz); 5.31 (1H, bs); 4.99(1H, bs); 4.43 (1H, m); 4.21 (1H, m); 2.57 (1H, dd, J=13.2, 3.62 Hz);2.36 (1H, dd, J=13.8, 5.8 Hz); 2.26 (1H, dd, J=13.4, 7.2 Hz); 1.95 (2H,m); 1.70-1.44 (12H, m); 1.42-1.36 (2H, m); 1.27-1.14 (2H, m); 1.20 (6H,s); 1.05-0.87 (2H, m); 0.23 (3H, s); 0.62 (3H, s).

EXAMPLE 109

Synthesis of Analogue 30

As described for 11.

Rf: 0.29 (dichloromethane:methanol 13:1).

IR (film): 3370 (s); 3082, 3045, 2964 (s), 1602, 1581, 1460, 1291 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 7.20 (1H, t, 7.96 Hz); 6.90 (1H, ddd, 0.8,1.5, 8 Hz); 6.86 (1H, dd, 1.6, 2.2 Hz); 6.71 (1H, ddd, 0.7, 2.1, 8 Hz);6.43 (1H, dd, 10.7, 15.5 Hz); 6.08 (1H, d, 10.7 Hz); 5.89 (1H, d, 15.5Hz); 5.31 (1H, dd, 1.4, 1.8 Hz); 5.00 (1H, br s); 4.44 (1H, dd, 5, 7Hz); 4.23 (1H, m); 3.96 (2H, t, 6.4 Hz); 2.57 (1H, dd, 3.73, 13.4 Hz);2.28 (1H, dd, 6.9, 13.4 Hz); 1.978 (1H, dd, 5, 7 Hz); 1.962 (1H, ddd,0.6, 4, 8 Hz); 1.50 (4H, q, 7.52 Hz); 1.39 (6H, S); 0.88 (6H, t, 7.52Hz).

EXAMPLE 110

Synthesis of Analogue 31

After protection of the tertiary alcohol as trimethylsilyl ether theappendage of the nor A-ring is done as usual. After removal of the silylether protective groups (TBAF, THF) the mixture is purified by columnchromatography (silica gel; dichloromethane:methanol 24:1) leading to amixture of the E-analogue 31 and its Z-isomer at 7,8 (ratio 2:1).

Rf: 0.20 (CH₃OH:CH₂Cl₂ 1:19).

¹H NMR: E-isomer (CDCl₃): δ 6.25 (1H, d, J=11.2); 5.94 (1H, d, J=11.3);4.10 (1H, m); 4.05 (1H, m); 2.81 (1H, m); 2.69 (1H, dd, J=3.8, 13.2);2.25 (1H, dd, J=7.8, 13.3); 1.20 (6H, s); 0.67 (3H, s).

¹H NMR: Z-isomer (CDCl₃): δ6.22 (1H, d, J=11.1); 6.08 (1H, d, J=11.1);4.10 (1H, m); 4.05 (1H, m); 2.39 (1H, dd, J=6.7, 13.4); 1.20 (6H, s);0.67 (3H, s).

EXAMPLE 111

Synthesis of Analog 32

As described for 11. Obtained together with the 7-Z-isomer (1:1).

Rf: 0.43 (dichloromethane:methanol 94:6).

IR (film): 3356-2924; 1436; 1374, 1205; 1144; 1054 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.30-6.29 (1H, 2×d, J=11.35, 11.11);6.17-6.14 (1H, 2×d, J=11.51-11.46); 5.48 (1H, m); 5.33 (1H, m); 5.00(1H, dpp s); 4.43 (1H, m); 4.22 (1H, m); 3.94 (1H, m); 2.60 (1H, d m,J=13); 2.35-2.15 (4H, m); 2.50-2.36 (2H, m); 1.23 (6H, 2×s); 2.12-1.91(5H, m); 1.88-1.79 (2H, m); 1.71-1.45 (7H, m) ppm.

EXAMPLE 112

Synthesis of Analogue 33

As described for 11. Obtained together with the 7-Z-isomer (6:4).

Rf: 0.31 (dichloromethane:methanol 94:6).

IR (film): 3367, 2936, 2865, 1433, 1363, 1308, 1217, 1152 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.25 (1H, 2×d, J=11.55, 11.65); 6.18 (1H,2×d, J=11.37, 11.36); 5.32 (1H, b s); 4.99 (1H, b s); 4.43 (1H, m; 4.21(1H, m); 3.99 (1H, ddd, J=7.2, 5.1, 5.1); 3.93 (1H, ddd, J=5.7, 4.9,4.9); 3.85 (1H, m); 3.77 (1H, m); 2.60 (1H, dd, J=13.18, 3.79); 2;49(1H, dd, J=6.0, 14.1); 2.45-1.23 (20H, m); 1.20 (6H, 2×s) ppm.

EXAMPLE 113

Synthesis of Analogue 34

As described for 11. Obtained together with the 7-Z-isomer (6:4).

Rf: 0.3 (dichloromethane:methanol 94:6).

IR (film): 3371, 2929, 2865, 1428, 1360, 1298, 1152, 1049, 974 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.18 (1H, 2×d, J=11.36, 11.46); 6.08 (1H,2×d, J=11.35, 11.5); 5.65 (2H, m); 4.10 (2H, m); 4.05 (H, m); 3.95 (1H,m); 3.90 (1H, m); 3.83 (1H, m); 2.71 (1H, dd, J=13.25, 3.75); 2.59 (1H,dd, J=13.6, 3.6); 2.52-1.56 (17H, m); 1.31 (3H, s); 1.29 (3H, s); 1.21(1H, m) ppm.

EXAMPLE 114

Synthesis of Analogue 35

As described for 11. Obtained together with the 7-Z-isomer (1:1).

Rf: 0.30 (dichloromethane:methanol 94:6).

¹H NMR: (500 MHz, CDCl₃): δ: 6.21 (1H, 2×d, J=9.32, 9.7); 6.08 (1H, 2×d,J=11.1, 11.71); 4.2-4.0 (4H, m); 2.62 (1H, d m, J=11.7); 2.49 (1H, dm,J=16.2) ppm.

EXAMPLE 115

Synthesis of Analogue 36

As described for 11. Obtained together with the 7-Z-isomer (1:1).

Rf: 0.26 (dichloromethane:methanol 94:6).

IR (film) 3390, 2925, 2855, 1458, 1361, 1172, 1051 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.28 (1H, 2×d, J=11.22, 11.38 Hz); 6.18(1H, 2×d, J=11.95, 11.17 Hz); 5.33 (1H, appd, J=1.45 Hz); 5.0 (1H, m);4.44 (1H, m); 4.21 (1H, m); 4.13 (1H, m); 4.04 (1H, m); 1.60 (1H, dm,J=12.80 Hz); 2.40 (5H, m); 2.20 (5H, m); 2.10 (4H, m); 1.92 (4H, m);1.85 (12H, m); 1.40 (12H, m); 1.25 (6H, 2×s); 1.21 (6H, 2×s).

EXAMPLE 116

Synthesis of the Analogue 37

From 11.19 as described for analogue 11.

Rf: 0.48 (CH₂Cl₂:MeOH 9:1).

IR (CH₂Cl₂): 3343 (br, s); 2962 (s); 2861 (m); 1640 (w); 1558 (w); 1456(s); 1375 (s); 1261 (m); 1057 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.34 (1H, dd, J=10.8, 15.1); 6.08 (1H, d,J=10.8); 5.65 (1H, dd, J=15.1, 8.6); 5.32 (1H, d, J=1); 5.01 (1H,d);4.42 (1H, m); 4.21 (12H, m); 2.58 (1H, dd, J=13.1, 3.9); 2.47 (1H);2.27 (1H, dd, J=7.6); 1.99 (1H, m); 1.95 (1H, m) 1.76 (1H, m); 1.60-1.20(14H, m); 1.21 (6H, s); 0.84 (3H, s); 0.76 (3H, s); 0.67 (3H, s) ppm

EXAMPLE 117

Synthesis of the Analogue 38

From 11.20 as described for analogue 11.

Rf: 0.19 (CH₂Cl₂:MeOH 95:5).

IR (CH₂Cl₂): 3380 (s); 2960 (s); 2939 (s); 2872 (m); 1633 (m); 1454 (s);1374 (s); 1253 (m); 1092 (s); 1054 (s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.34 (1H, dd, J 10.8, 15.1); 6.08 (1H, d);5.65 (1H, dd, J=8.5); 5.32 (1H, d, J=1); 5.01 (1H, d); 4.42 (1H, m);4.22 (1H, m); 2.58 (1H, dd, J=13.2, 4.0); 2.47 (1H, dd); 2.27 (1H, dd,J=7.6); 2.23-1;90 (2H, m); 1.76 (1H, m); 1.60-1.50 (5H, m); 1.46 (4H, q,J=7.5); 1.50-1.38 (4H, m); 1.35-1.15 (5H, m); 0.86 (6H, t); 0.84 (3H,s); 0.76 (3H, s); 0.67 (3H, s) ppm.

EXAMPLE 118

Synthesis of the Analogue 39

From 11.21 as described for analogue 11.

Rf: 0.20 (CH₂Cl₂:MeOH 95:5).

IR (CH₂Cl₂): 3402 (s); 2967 (s) 2872 (m); 1634 (w); 1422 (m); 1373 (m);1265 (s); 1138 (w) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.35 (1H, dd, J=10.8, 15.1); 6.21 (1H, dd,J=10.8, 15.5); 6.07 (1H, d); 5.96 (1H, dd, J=15.5); 5.75 (2H, 2×d); 5.63(1H, dd, J=8.6); 5.31 (1H, d, J=<1); 5.01 (1H, d); 4.44 (1H, m); 4.22(1H, m); 2.57 (1H, dd); 2.50 (1H, dd, J=8.9); 2.26 (1H, dd); 2.02-1.83(4H, m); 1.55 (m); 1.45 (m); 1.34 (6H, s); 0.98 (3H, s); 0.77 (3H, s);0.65 (3H, s) ppm.

EXAMPLE 119

Synthesis of the Analogue 40

From 11.27 as described for analogue 11. Also the 7,8-Z-isomer 40Z isformed (ratio 34:34′ 4:1). They can be separated y column chromatographyon silver nitrate impregnated silica gel (eluens MeOH:CH2Cl2 1:24→1:6).

40: Rf: 0.14 (MeOH:CH₂Cl₂ 1:14 on AgNO₃-silica gel).

¹H NMR: (500 MHz, CDCl₃): δ: 6.30 (1H, dd, J=10.8, 15.2); 6.08 (1H, d,J=10.8); 5.60 (1H, dd, J=8.8, 15.2); 5.30 (1H, br s); 4.98 (1H, d,J=1.8); 4.43 (1H, m); 4.20 (1H, m); 2.58 (1H, dd, J=4.0, 13.0); 2.3 (1H,q, J=9); 2.25 (1H, J=8.3, 13.0); 2.05 (1H, m); 1.88 (1H, ddd, J=3.8.8.3, 13); 1.78 (1H, m); 1.21 (6H, s); 0.93 (2H, t, J=7); 0.94 (3H, d,J=7.0); 0.85 (3H, t, J=6.6); 0.65 (3H, s) ppm.

40Z: Rf: 0.10 (MeOH:CH₂Cl₂ 1:14 on AgNO₃-silica gel).

¹H NMR: (500 MHz, CDCl₃): δ: 6.35 (1H, t, J=11); 6.26 (1H, d, J=12.5);5.33 (1H, dd, J=1, 2); 5.28 (1H, t, J=11); 5.01 (1H, br s);4.43 (1H, m);4.22 (1H, m); 2.84 (1H, q, J=9); 2.59 (1H, dd, J=4.1, 13.2); 2.31 (1H,dd, J=6.9, 13.4); 1.97 (1H, ddd, J=4, 8, 1.2); 1.82 (1H, m); 1.22 (6H,s); 0.96 (3H, d, J=6.7); 0;94 (3H, t, J=7.3); 0.84 (2H, t, J=6.5); 0.70(3H, s) ppm.

EXAMPLE 120

Synthesis of Analogue 41

As described for 11.

Rf: 0.37 (dichloromethane:methanol 9:1).

IR (film): 3376 (s, br), 2934 (s), 2242 (w), 1631 (w), 1461 (s), 1381(m), 1056 (s), 958 (m), 911 (s) cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 6.41-6.30 (1H, m); 6.10-6.00 (1H, m);5.70-5.59 (1H, m); 5.31 (1H, d); 5.00 (1H, s, br); 4.44 (1H, m); 4.22(1H, m);2.60-2.52 (1H, m); 2.30-2.00 (4H, m); 1.96 (2H, t); 1.90-1.10(14H, m); 1.05 (6H, t); 0.95-0.80 (6H, m).

EXAMPLE 121

Synthesis of Analogue 42

As described for 11

Rf: 0.28 (dichloromethane:methanol 9:1).

IR (film): 3382 (s); 2925, 1660, 1455, 1261, 1055 cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 7.12 (1H, dd, 11.2, 15.5 Hz); 6.30 (1H, d,15.5 Hz); 6.19 (1H, d, 3.3 Hz); 6.18 (1H, d, 11 Hz); 6.17 (1H, d, 3.3Hz); 6.06 (1H, dt, 1, 11 Hz); 5.50 (1H, ddd, 7, 8, 11 Hz); 5.27 (1H, d,2 Hz); 5.05 (1H, d, 2 Hz); 4.42 (1H, m, W1/2 11 Hz); 4.25 (1H, m, W1/219 Hz); 2.91 (1H, m); 2.64 (1H, dm, 13 Hz), 2.44 (2H, m); 2.31 (1H, dd,8.5, 13 Hz); 1.83 (1H, ddd, 4, 9, 13 Hz); 0.88 (3H, t, 7.5 Hz); 0.86(3H, t, 8 Hz).

EXAMPLE 122

Synthesis of Analogue 43

As described for 13.

Rf: 0.39 (dichloromethane:methanol 9:1).

IR (film): 3377 (s, br), 2931 (s), 1610 (w), 1454 (s), 1376 (s), 1265(s), 1214 (w), 1152 (w), 1049 (s), 976 (m) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.26 (1H, d, J=11.2); 6.04 (1H, d); 4.09(2H, m); 2.69 (1H, dd, J=3.8, 13.3 Hz); 2.47 (2H, m); 2.29 (1H, dd,J=13.3, 7.8 Hz); 2.21-2.07 (3H, m); 1.91 (1H, m); 1.85 (2H, m);1.73-1.30 (17H, m); 1.23 (6H, s); 1.05 (1H, m); 0.93 (3H, s); 0.88 (3H,d, J=6.6 Hz);

EXAMPLE 123

Synthesis of Analogue 44

As described for 13.

Rf: 0.062 (ethyl acetate:hexane 5:95).

IR (film): 3379, 2927, 2291, 3224, 1608, 1452, 1374, 1261, 1125, 1087.3,1044 cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 4.10 (2H, dddd); 6.25 (1H, d, J=11.3 Hz);6.05 (1H, d, J=11.4 Hz); 0.95 (3H, s); 1.03 (3H, d, J=6.49 Hz); 1.50(6H, s); 2.49 (1H, dd, J=13.4, 3.55 Hz); 2.69 (1H, dd, J=13.3, 3.83 Hz).

EXAMPLE 124

Synthesis of Analogue 45

As described for 13.

Rf: 0.24 (dichloromethane:methanol 4:96).

IR (film): 3422, 2976, 1642, 1451, 1267, 1088, 1048, 880 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ:: 1.53 (3H, s); 2.97 (1H, d, J=5.55 Hz);2.73 (1H, d, J=5.54 Hz); 1.04 (3H, d, J=5.17 Hz); 0.95 (3H, s); 6.25(1H, d, J=11.38 Hz); 6.04 (1H, d, J=11.30 Hz); 4.00 (2H, m).

EXAMPLE 125

Synthesis of Analogue 46

As described for 13.

Rf: 0.17 (dichloromethane:methanol 95:5).

IR (film): 3360, 3040, 2233, 1647, 1611, 811 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 0.93 (3H, s); 0.94 (3H, d, J=7.81 Hz); 1.49(6H, s); 4.09 (2H, m); 6.04 (1H, d, J=11.11 Hz); 6.25 (1H, d, J=11.11Hz).

EXAMPLE 126

Synthesis of the Analogue 47

From 12.14 as described for analogue 13.

Rf: 0.24 (CH₂Cl₂:MeOH 95:5).

UV (MEOH) λ_(max)=249 nm.

IR (film): 3360; 3036; 1649; 1610; 811 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.26 (1H, d, J=11.31); 6.03 (1H, d,J=11.31); 4.09 (2H₁ m); 1.21 (6H, s); 0.92 (3H, d) 0.84 (3H, d, J=6.61)ppm.

MS: m/z 386 (3); 353 (1; 303 (1); 45 (100).

EXAMPLE 127

Synthesis of the Analogue 48

From 12.13 as described for 13.

Rf: 0.31 (CH₂Cl₂:MeOH 95:5).

IR (film): 3361; 3036; 2236; 1612; 811 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.25 (1H, d, J=1t04); 6.04 (1H, d, J=11.04Hz); 4.10 (2H, m); 1.02 (6H, t, J=7.37); 0.95 (3H, d, J=6.71); 0.93 (3H,s) ppm.

EXAMPLE 128

Synthesis of the Analogue 49

From 12.15 and 13.2 as described for 19 from 6.26.

Rf: 0.17 (CH₂Cl₂:MeOH 95:5).

IR (film): 3364; 3026; 1599; 990; 810 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.26 (1H, d, J=11.20); 6.16 (1H, dd,J=15.44, 0.30); 6.04 (1H, d, J=11.20 ); 5.95 (1H, dd, J=15.29, 10.30);5.70 (1H, d, J=15.44); 5.58 (1H, dd, J=15.28, 8.22); 4.08 (2H, m); 1.34(6H, s); 0.96 (3H, d, J=6.70); 0.93 (3H, s) ppm.

EXAMPLE 129

Synthesis of the Analogue 50

From 12.15 as described for 20 from 6.26.

Rf: 0.26 (CH₂Cl₂:MeOH ˜95:5).

¹H NMR: (500 MHz, CDCl₃): δ: 6.26 (1H, d, J=11.16); 6.16 (1H, dd,J=15.46, 10.32); 6.04 (12H, d, J=11.16); 5.97 (1H, d,d J=15.24, 10.32);5.57 (1H, dd, J=15.24, 8.07); 5.52 (1H, d, J=15.46); 4.09 (2H, m); 0.98(3H, d, J=6.71); 0.94 (3H, s); 0.87 (6H, dd, J=7.44, 7.44) ppm.

EXAMPLE 130

Synthesis of the Analogue 51

From 12.16 as described for 19 from 6.26.

Rf: 0.17 (CH₂Cl₂:MeOH ˜95:5).

IR (film): 3360; 3036; 1610; 811 cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 6.26 (1H, d, J=11.09); 6.03 (1H, d,J=11.09); 4.08 (2H, m); 1.20 (6H, s); 0.92 (3H, s); 0.82 (3H, d, J=6.48)ppm.

MS: m/z 400 (29); 382 (10); 303 (4); 275 (11); 257 (12); 59 (100).

EXAMPLE 131

Synthesis of the Analogue 52

From 12.16 as described for 20 from 6.26.

Rf: 0.28 (CH₂Cl₂:MeOH ˜95:5).

IR (film) 3364; 3037; 1611; 811 cm⁻¹.

¹H NMR: (360 MHz, CDCl₃): δ: 6.26 (1H, d, J=10.98); 6.03 (1H, d,J=10.98); 4.08 (2H, m); 1.45 (4H, q, J=7.40); 0.92 (3H, s); 0.85 (6H, t,J=7.40); 0.82 (3H, d, J=6.55) ppm.

MS: m/z428 (34); 381 (11); 299 (6); 45 (100).

EXAMPLE 132

Synthesis of the Analogue 53

From 12.17 as described for 19 from 6.26.

Rf: 0.20 (CH₂Cl₂:MeOH ˜95:5).

IR (film): 3370 (s, br); 3051 (w); 2970 (m); 1607 (m); 1263 (s); 1095(s) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.23 (2H, m); 5.99 (2H, m); 5.73 (1H, d,J=15.5): 5.65 (1H, dd, J=8.5, 15.35); 4.10 (1H, m); 4.06 (1H, m); 2.69(1H, m); 2.48 (3H, m); 2.26 (1H, dd, J=5.0, 13.0); 2.15 (2H, m); 1.91(1H, m); 1.83 (3H, m); 1.72 (1H, m); 1.62-1.44 (4H, m); 1.35 (6H, 2s);1.25 (3H, m); 0.99 (3H, d, J=6.8); 0.94 (3H, s) ppm.

EXAMPLE 133

Synthesis of the Analogue 54

From 12.18 as described for 19 from 6.26.

Rf: 0.19 (CH₂Cl₂:MeOH ˜95:5).

IR (CH₂Cl₂): 3372 (s); 2928 (s); 1620 (w); 1462 (m); 1374 (s); 1019 (s);974 (m) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.26 (1H, d, J=11.3); 6.04 (1H, d. J=11.3);4.09 (2H, m); 2.69 (1H, dd, J=3.8, 13.2); 2.47 (3H, m); 2.30 (1H, dd,J=7.7, 13.3); 2.19 (1H, dd, J=16.4, 13.2); 2.10 (1H, m); 1.90 (1H, m);1.84 (2H, m); 1.70 (1H, m); 1.62-1.23 (19H, m); 1.22 (6H, s); 0.93 (3H,s); 0.87 (3H, d, J=6.6) ppm.

MS: m/z

EXAMPLE 134

Synthesis of the Analogue 55

From 12.18 as described for 20 from 6.26.

Rf: 0.19 (CH₂Cl₂:MeOH ˜95:5).

IR (film): 3383 (s, br); 2927 (s); 161 0 (m); 1046 (s); 974 (m) cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 6.26 (1H, d, J=11.2); 6.04 (1H, d, J=11.2);2.46 (2H, m); 2.29 (1H, dd, J=7.7, 13.2); 2.19 (1H, dd, J=6.5, 13.3);2.10 (2H, m); 1.91 (1H, m); 1.84 (2H, m); 1.70 (1H, m); 1.62-1.50 (6H,m); 1.47 (4H, q, J=7.5); 1.50-1.17 (13H, m); 0.92 (3H, s); 0.86 (6H, t,J=7.5); 0.86 (3H, d, J=6.5) ppm.

EXAMPLE 135

Synthesis of the Analogue 56

From 10.8 as described for 19 from 6.26.

Rf: 0.26 (hexane:acetone 6:4)

IR (film): 3388, 2927, 1634, 1464, 1367, 1058, 909, 734 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: :6.31 (1H, dd, J=15.16, 10.81); 6.05 (1H,d, J=10.84); 5.65 (1H, dd, J=15.15, 8.97); 5.30 (1H, m); 4.99 (1H, m);4.43 (1H, m); 4.22 (1H, m); 2.58 (1H, dd, J=13.32, 3.88); 2.16 (1H, dd,J=13.15, 7.11); 2.00 (1H, m); 1.94 (1H, m); 1.77-1.71 (3H, m); 1.54-1.24(m); 1.21 (6H, s); 0.896 (3H, d, J=7.58); 0.889 (3H, s); 0.740 (3H, s)ppm.

EXAMPLE 136

Synthesis of the Analogue 57

From 10.9 as described for 19 from 6.26.

Rf: 0.26 (hexane:acetone 6:4)

IR (film): 3387, 2934, 2865, 1634, 1454, 1366, 1057, 736 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: :6.34 (1H, dd, J=14.98, 10.65); 6.07 (1H,d, J=10.86); 6.05 (1H, dd, J=15.11, 9.62); 5.31 (1H, m); 4.99 (1H, m);4.44 (1H, m); 4.23 (1H, m); 2.57 (1H, d, J=13.14, 3.78); 2.28 (1H, dd,J=13.18, 6.69); 1.97 (2H, m); 1.88 (1H, m); 1.78 (1H, m); 1.65 (1H, m);1.53-1.24 (m); 1.21 (6H, s); 0.958 (3H, s); 0.906 (3H, d); 0.830 (3H, s)ppm.

EXAMPLE 137

Synthesis of 16.3

A suspension of (−)-quinic acid (16.1 47.5 g, 0.24 mol) and TsOH (200mg) in toluene (400 ml) is refluxed and the H₂O formed is removed with aDean-Stark apparatus. After 12 h, the mixture is filtered and dried(Na₂SO₄). Solvent evaporation gives crude 16.2 (42 g, 99%) which is usedas such in the next step.

A mixture of 16.2 (1.1 g, 6.3 mmol), t-butyldimethylsilyl chloride (1.09g, 7.24 mmol), DMAP (13 mg, 0.11 mmol) and imidazole (549 mg, 8.08 mmol)in DMF (5.8 ml) is stirred for 12 h at r.t. under nitrogen. The mixtureis diluted with Et₂O, quenched with H₂O and extracted with Et₂O. Theorganic layer is washed with brine, dried (Na₂SO₄), filtered andconcentrated. Column chromatography (silicagel; hexane:EtOAc 2:1) andHPLC separation (CH₂Cl₂:MeOH, 97:3) gives 16.3 (1.16 g, 66%). M.p.94-96° C.

Rf: 0.29 (hexane:EtOAc 2:1).

IR (film): 3480, 3308, 1782, 1150, 1085 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.87 (1H, dd, J=4.9, 6.0); 3.97 (1H, dd,J=4.4, 4.9); 3.89 (1H, ddd, J=4.4, 7.0, 10.8); 2.97 (1H, s, D₂Oexchangeable), 2.79 (1H, s, D₂O exchangeable); 2.62 (1H, d, J=11.6);2.29 (1H, ddd, J=2.8, 6.0, 11.6); 2.02 (1H, ddd, J=2.8, 1.0, 12.1); 1.97(1H, dd, J=10.8, 12.1); 0.91 (9H, s);0.10 (6H, s) ppm.

EXAMPLE 138

Synthesis of 16.5

A mixture of 16.3 (8.43 g, 29.2 mmol), 1,1-thiocarbonyidiimidazole (28.3g, 0.154 mol) and DMAP (203 mg, 1.67 mmol) in dichloroethane (80 ml) isrefluxed for 3 days. The solution is decanted and the residue is washedwith warm CH₂Cl₂. Evaporation of the combined organic phases andchromatography (silicagel; hexane:EtOAc 1:4) gives 16.4 (12.9 g, 87%).Tributyltin hydride (0.42 ml, 1.58 mmol) is added dropwise to a solutionof 16.4 (200 mg, 0.395 mmol) and AlBN (8 mg) in degassed dry toluene (5ml). After reflux for 5 h, the solvent is evaporated. Columnchromatography (silicagel; hexane:EtOAc 5:1) gives 16.5 (56 mg, 55%).M.p. 52-54° C.

Rf: 0.60 (hexane:EtOAc 2:1).

IR (film): 1777, 1259, 1124, 838, 776 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.84 (1H, dd, J=5.7, 5.0); 4.03 (1H, ddd,J=6.5, 6.4, 9.6, 9.6); 2.68 (1H, m); 2.41 (1H, ddddd, J=1.9, 2.0, 5.0,6.5, 13.4); 2.35 (1H, dddd, J=1.9,2.0, 5.7; 11.5); 2.24 (1H, ddddd,J=2.0, 2.0, 4.9, 6.4, 12.7); 1.81 (1H, d, J=11.5); 1.58 (1H, m); 1.52(1H, dd, J=9.6, 13.4); 0.88 (9H, s); 0.05 (6H, s) ppm.

EXAMPLE 139

Synthesis of 16.6

A 30% solution of NaOMe in dry MeOH (3.7 ml, 19.47 mmol) is added to16.5 (2.49 g, 9.73 mmol) in dry MeOH (40 ml) at 0° C. under nitrogen.After stirring for 1 h at 0° C., saturated NH₄Cl solution (40 ml) isadded and the solution is neutralized with 2 N HCl. The mixture isextracted with CH₂Cl₂, the combined organic layer is washed with brine,dried (MgSO₄). Filtration, solvent evaporation and filtration over ashort pad of silicagel (hexane:EtOAc 2:1) gives pure 16.6 (2.8 9, 100%).

Rf: 0.28 (hexane:EtOAc 2:1).

IR (film): 3385, 1739, 1257, 1039, 837, 778 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.24 (1H, s); 4.04 (1H, m); 3.70 (3H, s);2.85 (1H, dddd, J=3.67, 3.67, 11.95, 11.95); 2.22 (1H, m); 1.98 (1H, m);1.85 (1H, m); 1.32-1.57 (3H, m); 0.90 (9H, s); 0.08 (6H, s) ppm.

EXAMPLE 140

Synthesis of 16.7

A mixture of 16.6 (2.77 g, 9.63 mmol), p-bromophenyl sulphonyl chloride(4.00 g, 15.6 mmol), DMAP (30 mg, 0.25 mmol) in anhydrous pyridine (4.6ml) and chloroform (1.8 ml) is stirred for 1.5 h at 0° C., and 12 h atr.t. Water and ether is added. The mixture is extracted with ether. Thecombined organic phase is washed successively with 2% HCl solution,saturated NaHCO₃ solution and water and is dried (MgSO₄). Filtration,concentration and chromatography (silicagel; hexane:EtOAc 5:1) gives16.7 (4.88 g, 100%).

M.p. 62-64° C.

Rf: 0.57 (hexane:EtOAc 2:1).

IR (film): 1737, 1577, 1369, 1188, 1049, 967, 822 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 7.73 (4H, m); 4.72 (1H, dddd, J=4.52, 4.52,11.29, 11.29); 4.19 (1H, m); 3.69 (3H, s); 2.81 (1H, dddd, J=3.64, 3.64,12.47, 12.47); 2.31 (1H, m); 1.86 (1H, m); 1.60 (1H, m); 1.43-1.52 (3H,m); 0.83 (9H, s); 0.02 (3H, s); −0.02 (3H, s) ppm.

EXAMPLE 141

Synthesis of 16.8

To a stirred solution of 16.7 (4.64 g, 9.15 mmol) in anhydrous t-BuOH(30 ml) is added dropwise a 1 M solution of t-BuOK in t-BuOh (10.6 ml,10.6 mmol) at 50° C., under N₂. The resulting mixture is refluxed for 1h. saturated NH₄Cl solution (20 ml), brine (10 ml) and water (5 ml) areadded. The mixture is extracted with ether. The combined organic phaseis dried (MgSO₄), filtered, and the solvent is evaporated below 18° C.Chromatography (silicagel; ether:pentane 5:95) gives 16.8 (1.63 g, 71%).

Rf: 0.48 (hexane:EtOAc 5:1).

IR (film) 1727, 1371, 1256, 1114, 1097, 838 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 3.93 (1H, m); 3.66 (3H, s); 2.20 (1H, dd,J=7.2, 12.9); 2.14 (1H, dd, J=8.2, 12.9); 2.07 (1H, dd, J=7.1, 12.0);1.81 (1H, m); 1.77 (1H, m); 1.29 (1H, dd, J=5.0, 8.5); 0.87 (9H, s);0.67 (1H, dd, J=5.0, 5.0); 0.02 (6H, s).

MS: m/z 239 (10, 213 (100), 199 (9), 167 (35), 149 (39), 125 (20), 111(18), 89 (96), 45 (98) ppm.

EXAMPLE 142

Synthesis of 16.9

To a stirred solution of 16.8 (570 mg, 2.11 mmol) in anhydrous toluene(25 ml) is added dropwise a solution of diisobutylaluminum hydride (5.28ml, 5.28 mmol) 1 M in hexane at −78° C., under N₂. Stirring is continuedfor 2 h at −78° C. The reaction is quenched with a 2 N solution ofpotassium sodium tartrate (25 ml). The stirring is continued overnightwhile the temperature gradually came to r.t. The mixture is extractedwith CH₂Cl₂, dried (MgSO₄) and evaporated. Chromatography (silicagel;hexane:EtOAc 4:1), purification gives 16.9 (500 mg, 98%).

Rf: 0.30 (hexane:EtOAc 4:1).

IR (film) 3328, 1256, 1115, 1094, 1032, 904, 775 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 4.03 (1H, m); 3.62 (1H, dd, J=5.1, 11.1);3.51 (1H, dd, J=5.1, 11.1); 2.05 (1H, dd, J=6.4, 12.6 Hz); 1.92 (1H, dd,J=6.4, 12.6); 1.75-1.84 (2H, m); 1.18 (1H, ddd, J=4.2, 4.2, 8.4); 0.89(9H, s); 0.51 (1H, dd, J=5.1, 8.4); 0.02 (6H, s); 0.38 (1H, dd, J=4.2,4.2) ppm.

EXAMPLE 143

Synthesis of 16.10

To a stirred solution of 16.9 (480 mg, 1.98 mmol) in dichloromethane (20ml) is added PCC (750 mg, 3.4 9 mmol) at r.t. under nitrogen. After 2 hstirring, the mixture is filtered over celite, which is washed withdichloromethane. The combined filtrate is washed successively withbrine, NaHCO₃ solution and brine. Drying (Na₂SO₄), filtration andchromatography (silicagel, ether:pentane 1:9) gives 16.10 (430 mg, 90%).

Rf: 0.40 (hexane:EtOAc 9:1).

IR (film): 1706, 1256, 1121, 1072, 838, 778 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 8.90 (1H, s); 4.04 (1H, m); 2.17 (1H, ddd,J=1.1, 8.0, 13.0); 2.13 (1H, dd, J=7.2, 13.0); 2.10 (1H, dd, J=7.2,13.0); 1.93 (1H, ddd, J=5.1, 5.3, 8.8); 1.80 (1H, ddd, J=5.1, 8.0,13.0); 1.35 (1H, dd, J=5.6, 8.8); 0.97 (1H, dd, J=5.3, 5.6); 0.88 (9H,s);0.04 (6H, s) ppm.

EXAMPLE 144

Synthesis of 16.11

To a suspension of t-BuOK (352 mg, 3.14 mmol) in dry THF (2 ml) is addeddropwise a solution of dimethyl diazomethyl phosphonate (219 mg, 1.45mmol) in dry THF (2 ml) at −78° C., under nitrogen. After 10 min, asolution of 16.10 (290 mg, 1.21 mmol) in dry THF (2 ml) is addeddropwise at −78° C. Stirring is continued, at −78° C. for 4 hrs, at −15°C. for 8 hrs, and at r.t. for 5 hrs. Water is added, followed byextraction with dichloromethane and drying (MgSO₄). Filtration, solentevaporation below 18° C. and chromatography (silicagel:pentane, thenether:pentane 1:9) gives 16.11 (254 mg, 89%) as a colorless oil.

Rf: 0.69 (hexane:EtOAc 9:1).

IR (film): 3467, 3315, 2113, 1111, 1095, 836, 776 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 3.84 (1H, m); 2.28 (1H, J=7.1, 12.5); 2.05(1H, dd, J=7.1, 12.7); 1.92 (1H, s); 1.90 (1H, ddd, J=1.0, 8.3, 12.5);1.83 (1H, ddd, J=4.9, 8.1, 12.7); 1.60 (1H, ddd, J=4.9, 4.9, 8.3); 0.88(9H, s); 0.81 (1H, dd, J=4.9, 8.1 Hz); 0.55 (1H, dd, J=4.9, 4.9); 0.01(6H, s) ppm.

EXAMPLE 145

Synthesis of 16.12

Sodium hexamethyldisilazide (1M in THF, 5.2 ml, 5.2 mmol) is added to(bromomethylene)triphenyl phosphonium bromide (2.35 g, 5.4 mmol) in dryTHF (7 ml) at −68° C. After 1 h, a solution of 12.2b (357 mg, 0.91 mmol)in 2 ml of THF is added. After stirring for 1 h, the mixture is allowedto reach r.t. and is stirred overnight. Filtration through a short padof celite, washing with hexane and concentration affords an oily residuewhich is chromatographed (silica gel, hexane) to provide (E)- and(Z)-16.12 (in 3:1 ratio) in a combined yield of 56% (237 mg).

Rf: 0.41 (hexane).

IR (film): 2955, 2874, 1622, 1462, 1380, 1235, 1043, 743 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 5.87 (1H, br s); 2.50 (1H, ddd, J=4.6, 4.8,14.5); 2.22 (1H, dd, J=8.1, 9.7); 2.10 (1H, m); 1.86 (1H, m); 1.28 (3H,s); 1.20 (6H, s); 0.94 (9H, t, J=8.0); 0.87 (3H, d, J=5.5); 0.56 (6H, q,J=8.0) ppm.

MS m/z: 115 (17); 103 (89).

EXAMPLE 146

Synthesis of 16.15

To a stirred solution of 16.12 (51 mg, 0.11 mmol) in ether (0.6 ml) isadded dropwise a solution of t-butyllithium (1.7 M in n-pentane, 0.16ml, 0.27 mmol) at −78° C., under argon and stirring is continued for 50min. Then a solution of 16.10 (12 mg, 0.05 mmol), in diethyl ether (0.2ml) is dropwise added. The mixture is stirred 1 h at −78° C. and isquenched with saturated aqueous NH₄Cl (2 ml) and extracted with Et₂O andEtOAc. The combined organic phase is dried (MgSO₄), concentrated,filtered over a short silica gel pad (EtOAc:hexane 1:6) and purified byHPLC (silica gel; EtOAc:hexane 1:9) to give an epimeric mixture (in aratio of 6:4) of (E)-16.15 (11 mg) and (Z)-16.15 (3.5 mg) in a combinedyield of 46%.

Rf: 0.59 (EtOAc:hexane 1:6).

IR (film) 3394, 2954, 2876, 1465, 1390, 1383, 1092, 1043 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 5.14 (1H, 2×d, J=8.5, 8.7); 4.42 (1H, d,J=7.7, min); 4.23 (1H, d, J=8.5, maj.); 4.02 (1H, m); 1.20 (3H, s); 0.94(12H, t, J=8.0, superposed with s); 0.87 (12H, s+d, superposed); 0.56(6H, q, J=8.0); 0.28 (1H, dd, J=4.4, 4.6, min); 0.23 (1H, dd, J=4.4, 4.6maj.); 0.02 (6H, 2×s); ppm.

EXAMPLE 147

Synthesis of 16.13

To a stirred solution of 16.11 (22 mg, 0.093 mmol) in dry THF (4 ml) at−50° C., n-butyllithium (1.6 M solution in n-hexane, 0.14 ml, 0.23mmol), is added. After stirring for 1 h 12.2b (40 mg, 0.10 mmol) in dryTHF (1 ml) is added. The temperature was allowed to reach r.t. andstirring was continued for 30 min. Quenching with water, extraction withEt₂O , usual work-up and HPLC purification (silica gel; EtOAc:hexane1:20) provide 16.13 (22 mg, 55% based on the recovered 12.2, 15 mg) as asingle diastereomer.

Rf: 0.52 (EtOAc:hexane 1:9).

IR (film): 3477, 2953, 2875, 2226, 1463, 1380, 1253, 1093 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 3.83 (1H, J=7.7, 7.8, 15.2); 2.23 (1H, dd,J=7.1, 12.5); 2.03 (1H, dd, J=7.1, 12.6); 1.20 (6H, s); 0.99 (3H, s);0.94 (9H, t, J=8.0); 0.88 (3H, d, J=6.6); 0.85 (9H, s); 0.73 (1H, dd,5.0, 8.3); 0.56 (6H, q, d=8.0); 0.52 (1H, dd, J=4.9, 4.9); 0.0 (6H, s)ppm.

EXAMPLE 148

Synthesis of 16.14

Alcohol 16.13 (18 mg, 29 μmol) is refluxed in THF (5 ml) in the presenceof LiAlH₄ (4 mg) and sodium methoxide (4 mg). After 2 h, the mixture iscooled, quenched with saturated NH₄Cl and extracted with Et₂O. Usualwork-up followed by chromatographic purification (silica gel,EtOAc:hexane 1:25) gives 16.14 (9 mg, 50%).

Rf: 0.43 (EtOAc:hexane 1:9).

IR (film) 3508, 2930, 2872, 2463, 1380, 1256, 1093, 837 cm⁻¹.

¹H NMR: (500 MHz, CDCl₃): δ: 5.34 (2H, s); 3.95 (1H, m); 2.06 (2H, m);1.27 (3H, s); 1.19 (6H, s); 0.94 (9H, t, J=8.0); 0.93 (3H, d, J=6.0);0.87 (9H, s); 0.55 (6H, q, J=8.0); 0.07 (1H, dd, J=3.2, 3.4); 0.02 (6H,2×s) ppm.

EXAMPLE 149

Synthesis of Analogue 43

a) From 16.15:

A mixture of (E)-16.15 (10 mg, 0.016 mmol), PTSA (0.9 mg), water (0.4ml) and 1,4-dioxane (1;5 ml) is stirred for 6 h at 63° C. The mixture istreated with sat. NaHCO₃ (1.5 ml) and extracted with CH₂Cl₂. Thecombined organic phase is dried (MgSO₄), concentrated, filtered over ashort silica gel column (acetone:hexane 4:6) and purified by HPLC(silica gel; MeOH:CH₂Cl₂ 5:95) giving 43 (5.0 mg, 78%).

b) From 16.14:

As described from 16.15; yield 40% next to the 7-Z-isomer (ratio 3:1).

What we claim is:
 1. A compound of the formula I:

in which P is hydrogen or alkyl; Y and Y′ are each hydrogen or, whentaken together, represent a group ═CH₂; W and W′ are each hydrogen; X isselected from the group consisting of a hydroxyalkyl, a hydroxyalkoxy,an alkoxy optionally comprising an epoxide function, a hydroxygen ahydroxyalkadiene, a hydroxyalkyne and isomeric forms thereof; and a)either R₁ and R₃ or R′₃ form a saturated 5- or 6-membered carbocyclicring and R₂, R′₂, R₃ or R′₃, R₄, R′₄, R₅ and R′₅ are each Independentlyhydrogen or alkyl; or b) R₂ or R′₂ and R₄ or R′₄ form a saturated 5- or6-membered carbocyclic ring, and R₁, R₂ or R′₂, R₃, R′₃, R₄ or R′₄, R₅and R′₅ are each independently hydrogen or alkyl; or c) R₃ or R′₃ and R₅or R′₅ form a saturated or unsaturated 5- or 6-membered carbocyclicring, and R₁, R₂, R′_(2,) R₃ or R′₃, R₄, R′₄, and R₅ or R′₅ are eachindependently hydrogen or alkyl; or d) R₃ or R′₃ taken at the same timewith R₁ and R₅ or R′₅ form a saturated 8- or 12-membered carbobicyclicring, and R₂, R′₂, R₃ or R′₃, R₄, R′₄, and R₅ or R′₅ are eachindependently hydrogen or alkyl.
 2. The compound according to claim 1,wherein P is hydrogen; Y and Y′, taken together, form a ═CH₂ group; R₁,R₂, R′₄, R₅ and R′₅ are each hydrogen; and X is a group hydroxyalkyne.3. The compound according to claim 2, wherein X is a group—CH₂—C≡C—C(C₂H₅)₂(OH).
 4. The compound according to claim 1, wherein R₁and R′₃ taken together form a carbocyclic C-ring as shown in IIIa1 andIIIa2, respectively, or a diastereoisomer of IIIa1 and IIIa2:

in which ring C may be saturated, unsaturated or substituted.
 5. Thecompound according to claim 1, wherein R₂ and R₄ taken together form acarbocyclic D-ring as shown in IIIb1 and IIIb2, respectively, or adiastereoisomer of IIIb1 and IIIb2:

in which ring D may be saturated, unsaturated or substituted.
 6. Thecompound according to claim 1, wherein R′₃ and R′₅ taken together form acarbocyclic E-ring as shown in IIIc1 and IIIc2, respectively, or adiastereoisomer of IIIc1 and IIIc2:

in which ring E may be saturated, unsaturated or substituted.
 7. Thecompound according to claim 1, wherein R′₃ taken together at the sametime with R₁ and R′₅ form a bicyclic CE-ring system as shown in IIIe1and IIIe2, respectively, or a diastereoisomer of IIIe1 and IIIe2:

in which: n is 1, 2, 3, or 4, and rings C and/or E is optionallysaturated, unsaturated or substituted.
 8. The compound according toclaim 7, wherein X is hydrogen, lower alkyl, hydroxy or a functionalgroup derived therefrom.
 9. The compound according to claim 4, havingone of the following structures:


10. The compound according to claim 5, having one of the followingstructures:


11. The compound according to claim 6, having one of the followingstructures:


12. A pharmaceutical preparation comprising a therapeutically effectiveamount of a compound of claim 1 and a pharmaceutically and/orveterinarily acceptable carrier or diluent.
 13. A pharmaceuticalpreparation comprising a therapeutically effective amount of a compoundof claim 4, and a pharmaceutically and/or veterinarily acceptablecarrier or diluent.
 14. A pharmaceutical preparation comprising atherapeutically effective amount of a compound of claim 3 and apharmaceutically and/or veterinarily acceptable carrier or diluent. 15.A pharmaceutical preparation comprising a therapeutically effectiveamount of a compound of claim 2 and a pharmaceutically and/orveterinarily acceptable carrier or diluent.